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  • 1992  (223)
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  • Genetics  (415)
  • 101
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. S199 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 102
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. S549 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 103
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 104
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. 61-80 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 105
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. 141-160 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 106
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. 181-200 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 107
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 108
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; yeast ; plasmid stability ; phosphoglycerate kinase ; carbon flux ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In this report the effects of phosphoglycerate kinase (PGK) overproduction on the physiology and plasmid stability in baker's yeast Saccharomyciae cerevisiae containing the PGK1 gene on an episomal plasmid are described. This examination reveals that there is a preferred intracellular level for this enzyme, amounting to 10-15% of the total soluble protein. Strains containing the plasmid and the host strain were grown in non-selective batch cultures and continuous culture, under different growth conditons. Plasmid-containing yeast strains stabilize the copy number of the episomal plasmid at a level at which the PGK concentration is about 12%. This stabilization is due to an equilibrium between normal plasmid loss and selective pressure because of advantages resulting from the increased amount of PGK under glucose-limited conditions. During respiro-fermentative growth, PGK-overproducing cells showed an increased respiration rate and decreased fermentative activity, compared to the host strain.The PGK1 gene can be applied as a direct positive selection marker to obtain a high episomal plasmid stability during growth on glucose. The results are consistent with previously reported data on the physiology and gene stability of PGK-overproducing yeast cells that contain multiple copies of the PGK1 gene integrated into the genome.
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  • 109
    ISSN: 0749-503X
    Keywords: Protein kinase ; chromosome III ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have sequenced a gene on chromosome III of Saccharomyces cerevisiae which codes for a putative serine/threonine protein kinase of 726 amino acids (calculated molecular weight 82 kDa). We have called this gene KIN82. The amino acid sequence of KIN82 is most similar to the cyclic nucleotide-dependent protein kinase subfamily and the protein kinase C subfamily. Gene disruption of KIN82 did not produce any phenotype when tested under a variety of conditons. Reduced stringency hybridizations revealed the presence of another genomic sequence with high homology to the carboxy-terminal catalytic domain of KIN82.
    Additional Material: 4 Ill.
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  • 110
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. 79-82 
    ISSN: 0749-503X
    Keywords: Site-directed mutagenesis ; Saccharomyces cerevisiae ; PCR ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have developed a simple procedure for the localized mutagenesis of yeast genes. In this technique the region of interest is first amplified under mutagenic chain reaction (PCR) conditions. Cotransformation of the PCR product with a gapped plasmid containing homology to both ends the PCR procuct allows in vivo recombination to repair the gap with the mutagenized DNA. This procedure is efficient, allows trageting of specific regions for mutagenesis, and requires no subcloning steps in Escherichia coli.
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  • 111
    ISSN: 0749-503X
    Keywords: Genome sequencing ; Saccharomyces cerevisiae ; chromosome XI ; BAF1 ; APN1 ; protein kinase ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report here the DNA sequence of a segment of chromosome XI of Saccharomyces cerevisiae extending over 10·7 kb. The sequence was determined using a double-strand sequencing strategy adapted from the random-clone strategy. The segment contaiins seven non-overlapping long open reading frames, YKL500, 505, 510, 513, 516, 518 and 520 and one tRNA gene. YKL505 and YKL513 are two already sequenced genes, the BAF1/ABF1 and the APN1 genes, respectively. YKL510 exhibits a strong homology to the RAD2 protein and YKL516 is presumabloy a protein kinase.
    Additional Material: 6 Ill.
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  • 112
    ISSN: 0749-503X
    Keywords: Yeast ; protein export ; heat shock ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A new secreted yeast glycoprotein with an Mr of about 400 kDa (gp400) has been found. The glycoprotein is an O-mannosylated oligomer, whose synthesis and export into culture medium are stimulated by heat shock. Intracellular transport of gp400 is carried out by membrane vesicles distinct form the known constitutive scretory vesicles. Immunological analysis revealed gp400 only in Saccharomyces species.
    Additional Material: 12 Ill.
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  • 113
    ISSN: 0749-503X
    Keywords: Chromosome III ; sequencing ; yeast ; Saccharomyces cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have determined the nucleotide sequence of a segment from chromosome III fo Saccharomyces cerevisiae extending over 7·9 kb between the PGK1 and CRY1 loci.The fragment contains seven open reading frames, YCR241, YCR242, YCR243, YCR244, YCR245, YCR246 and YCR247, of more than 70 codons. The study of the effects of a global disruption of YCR242, YCR243, YCR244, YCR245 and YCR247 shows that they are not essential for growth division.
    Additional Material: 4 Ill.
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  • 114
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. 241-241 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 115
    ISSN: 0749-503X
    Keywords: Candida maltosa ; automonous replicating sequence ; nucleotide sequence ; transformation ; RS15 protein ; intron ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A Candida maltosa chromosomal DNA fragment which confers high frequency transformation of C. maltosa and autonomous replication of recombinant plasmids was cloned and sequenced. Analysis of the nucleotide sequence of the cloned DNA revealed a sequence homologous for C. maltosa autonomously replicating sequence (ARS) elements. Vector pRJ1 for C. maltosa was constructed, which contained a 1.3 kb ARS sequence, pICEM-19H and the ADE1 gene of C. maltosa. Southern blot analysis suggested that the copy number of pRJ1 in C. maltosa was approximately 20 per genome. The sequence analysis also revealed an open reading frame, encoding a polypeptide with high homology (70%) to the RS15 protein of Brugia pagangi. This open reading frame has an intron with canonical sites for correct splicing in Saccharomyces cerevisiae.
    Additional Material: 5 Ill.
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  • 116
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. 311-314 
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; genetic mapping ; CAR1 ; arginase ; arginine catabolism ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Additional Material: 2 Ill.
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  • 117
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XI ; mitochondria ; G Protein ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have entirely sequaenced an 8.3 kb segment localized on the left arm of chromosome XI of Saccharomyces cerevisiae. Five new open reading frames have been uncovered. One of them encodes a new mitochondrial carrier protein which is dispensable for growth on glycerol medium. Another could be a new member of the G protein family. A third possesses the PAAKK motif common to H1 histones.
    Additional Material: 7 Ill.
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  • 118
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. 373-384 
    ISSN: 0749-503X
    Keywords: Yeast ; double-stranded RNA virus ; K28 killer virus ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The killer phenotype expressed by Saccharomyces cerevisiae strain 28 differs fron that of the more extensively studied K1 and K2 killers with respect to immunity, mode of toxin action and cell wall primary toxin receptor. We previosly demonstrated that the M28 and L28 dsRNAs found in strain 28 are present in virus-like particles (VLPs) and that transfection with these VLPs is sufficient to confer the complete K28 phenotype on a dsRNA-free recipient cell. We also demonstrated that L28, like the L-A-H species in K1 killers, has [HOK] activity required for maintenance of M1-dsRNA, and predicted that M28 would share with M1 dependence on L-A for replication. We now confirm this prediction by genetic and biochemical analysis of the effects of representative mak, ski and mkt mutations on M28 maintenance, demonstrating that M28 replication resebles M1 in all respects. We also show that L28 is an L-A-H species lacking [B] activity, and that M28 excludes both M1 and M2 from the same cytoplasm. Stable coexpression of K28 phenotype from M28 and of K1 phenotype from an M1-cDNA clone was demonstrated. Exclusion, therefore, acts at the level of dsRNA replication, presumably reflecting competition for the L-A-H encoded capsid and cap-pol fusion protein, rather than reflecting incompatibility of toxin or immunity expression. Finally, we show that expression of active K28 toxin, bu t not of K28 immunity, requires the Kex2 endoprotease.
    Additional Material: 5 Ill.
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  • 119
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have isolated and sequenced a novel gene from Saccharomyces cerevisiae coding for an essential protein of unknown function. The gene called YJU2 was borne on a chromosome X fragments shown by hybridization to intact S. cerevisiae chromosomal DNA fractionated by orthogonal pulsed field electrophoresis. Northern hybridization analysis indicated that the 0.8-kb transcript of YJU2 is expressed in exponential-phase cells grown in rich medium (data not shown). Figure 1 shows the nucleotide and deduced amino-acid sequences of the 834-bp coding region as well as the nucleotide sequences of the 5′ upstream region and of the 3′ downstream region, together with the flanking neighbouring open reading frames (ORFs), YJU1 and YJU3.
    Additional Material: 2 Ill.
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  • 120
    ISSN: 0749-503X
    Keywords: Omnipotent suppressor ; SUP35 ; SUP45 ; elongation factor ; overexpression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The overexpression of SUP35 (SUP2) wild-type gene, caused by increase of its copy number, induces an omnipotent suppression similar to the phenotype of mutants for this gene. The effectt of extra-SUP35 was detected for moderate or even low copy number. Moreover, overdosage of the fragment including only the 5′-flanking region and N-terminal 100 bp of protein-coding sequence of SUP35 leads to allosuppression. Multi-SUP35 gene was also icompatible with extrachromosomal suppressor factor ψ, presumbaly because of high level of mistranslation. The suppressor effect caused by overdosage of another gene, SUP45 (SUP1), is much lower and can be detected only for one construction which is derived from high copy number plasmid. Suppression induced by extra-SUP35 and especially by extra-SUP45 is affected by the cell environment. A model predicting that the balance of gene products is a key for regulation of translational fidelity is discussed.
    Additional Material: 4 Ill.
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  • 121
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. 559-567 
    ISSN: 0749-503X
    Keywords: MSS51 ; QRI5 ; chromosome XII: ABF1 binding site ; Saccharomyces cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have localized gene MSS51 on chromosome XII of Saccharomyces cerevisiae between the RDN1 and CDC42 loci. ‘Head to head’ with MSS51 is another gene, QRI5, the function of which is unkown. However, the proximity of these genes, the structure of the intergenic region and the presence of an ABFl binding site right in the middle of this region suggest that the MSS51 and QRI5 expressions are submitted to a common regulatory process.
    Additional Material: 5 Ill.
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  • 122
    Electronic Resource
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. ii 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 123
    ISSN: 0749-503X
    Keywords: Heterologous gene expression ; native proteins ; Saccharomyces cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A genetal method is described for screening Saccharomyces cerevisiae colonies for the intracellular expression of native proteins. Colonies are replicated onto nitrocellulose membranes and yeast walls are removed enzymatically. The resulting spheroplasts are rapidly lysed by placing chromatography paper soaked in hypotonic buffer on the membranes. Intracellular proteins released by spheroplast lysis are bound in situ to the nitrocllulose under non-denaturing conditions and potentially can be examined using enzymatic or immunologeic methods. For example, in the present study colonies were screened for the presence of elongation factor 2 (EF-2) that can be [32P]ADP-ribosylated by diphtheria toxin and [32P]NAD+. Recognition by the toxin requires the presence in EF-2 of the unique posttranslationally modifed histidine derivative, dipthamide. The procedure described here reliably discriminates between wild-type yeast colonies and mutant colonies that do not synthesize diphthamide. In addition to faciliating the study of dipthamide biosynthesis in yeast, the more general application of this procedure of this procedure will enable the screening of colonies with assays that require native proteins.
    Additional Material: 1 Ill.
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  • 124
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. 655-665 
    ISSN: 0749-503X
    Keywords: Maltose utilization genes ; Saccharomyces cerevisiae ; telmoeres ; repeated genes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Saccharomyces strains capable of fermenting maltose contain any one of five telomere-associated MAL loci. Each MAL locus is a complex of three genes encoding the three functions required to ferment maltos: maltose permease (GENE 1), maltase (GENE 2) and the MAL trans-activator (GENE 3). All five loci have been cloned and all are highly sequence homologous over least a 9-0 kbp region containing these GENEs (Charron et al., Genetics 122, 307-331, 1989). Our initial studies of strians carrying the MAL3 locus indicated the presence of linked, repeated MLA-homologous sequences (Michels and Needleman, Mol. Gen. Genet. 191, 225-230, 1983). Here we report our analysis of the centromere-proximal MAL3-linked sequence and show that the complete MAL3 locus spans approximately 40 kbp and consists of tandemly arrayed, partial repeats of the three GENE sequences described above. In addition, the structure of the MAL3 locus is compared to that of three partially functional alleles of MAL3. These alleles were shown to contain only MAL31 and MAL32 and their structure suggests that they resulted from MAL3 deletions removing the sequences centromere-proximal of MAL31. The amplification and rerrangement of the telomer-linked MAL3 sequences are discussed in the context of studies on other telemere-associated sequences from yeast and other species.
    Additional Material: 4 Ill.
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  • 125
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Additional Material: 4 Ill.
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  • 126
    Electronic Resource
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. 699-710 
    ISSN: 0749-503X
    Keywords: MOL1 ; molasses ; stationary phase ; stress ; tRNA-gly(UCC) ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have isolated a new Saccharamyces cerevisiae gene, MOL1, that is transiently expressed at high levels in the early stationary phase of batch cultures growing on industrial molasses medium. The DNA sequence of the M O L I gene (for MOLasses-inducible) with its flanking regions was determined (EMBL accession number X61669). It encodes a polypeptide of M, 35 kDa that is closely related to stress-inducible proteins of similar size frow two Fusarium species. Unlike ST135 of Fusarium, MOL1 is not induced by ethanol or heat shock. MOL1 expression is absent in rich (YP) medium, and only very low levels of expression are detectable in minimal (YNB) medium. The gene is not essential, and a MOL1 distruption strain showed no apparent phenotype undera variety of growth conditions.The 5′ region of MOL1 contains the complete sequence previously determined for the SUF4 locus, encoding a tRNA-gly (UCC) gene, which has been mapped to chromosome VII.
    Additional Material: 8 Ill.
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  • 127
    ISSN: 0749-503X
    Keywords: Isoprene ; chromosome ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Isopontenyl diphosphate isomerase catalyses an essential activation step in the isoprene biosynthetic pathwasy. The Saccharomyces cerevisiae gene for isomerase, IDI1, was recently isolated and characterized (Anderson et al. J. Biol. Chem. 1989aa, 264, 19169-19175). Wild-type IDI1 was disrupted with a LEU2 marker, and the resulting DNA was used to transform a yeast leucine auxotroph. Southern blots of EcoRI fragments of chromosomal DNA from the diploid strain showed the expected fragments for intact and disrupted IDI1. Dissection and analysis of tetrads demonstrated that IDI1 is an essential single-copy gene. A CHEF gel and clone grid filter analysis, followed by chromosomal mapping indicated that the gene is located on chromosome that the gene is located on the chromosome XVI approximately 55 kb contromere proximal to PEP4.
    Additional Material: 5 Ill.
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  • 128
    ISSN: 0749-503X
    Keywords: Yeast ; Saccharomyces cerevisiae ; Chromosome II ; tRNAGly ; tRNALeu ; sigma ; delta ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The DNA sequence of an 8079 bp ClaI fragment located at 40 kb from the centromere on the left arm of chromosome II from Saccharomyces cerevisiae has been determined. Sequence analysis reveals five new oen reading frames, tRNAGly and tRNALeu genes as well as sigma and truncated delta elements. The disruption of the three larger open reading frames shows that they are not essential for mitotic growth.
    Additional Material: 5 Ill.
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  • 129
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. 801-804 
    ISSN: 0749-503X
    Keywords: Kluyveromyces lactis ; LEU2 gene ; rat ribosomal protein L7 ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A DNA fragment that can complement the leu2 mutation of Saccharomyces cerevisiae was cloned from the genomic library of Kluyveromyces lactis.The nucleotide sequence revealed an open reading frame of 362 codons, 75% homologous to S. cerevisiae LEU2 gene. The upstream region contained a CCGGAACCGG sequence identical to the site of leucine-specific control of LEU2. Further upstream, there is a partial open reading frame homologous to rat ribosmal protien L7.
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  • 130
    Electronic Resource
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 131
    ISSN: 0749-503X
    Keywords: mitochondria ; immunogold labelling ; HSP70 ; in vitro import ; respiration ; Q pool ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A technique is described for the isolation and purification of intact, respiratory-competent mitochondria from Sachizo-saccharomyces pombe. The purified mitochondria are capabel of oxidizing NADH and succinate as resporatory substrates. indicating the presence of succinate dehydorgenase and an NADH dehydrogenase located on the outer suface of the inner membrane. Mitochondria display good respiratory control with an ADP/O ratio of 〈2. Respiratory activity is linearly dependent upon the redox poise of the quinone pool, suggesting the presence of an unbranched repiratory pathway to molecular oxygen. Immunogold labelling using antisera raised against mitochodria proteins (SSPI, SSCI, and PHSPI) from three different species, namely S. pombe, Saccharomyces cerevisiane and the plant Pisum sativum respectively, has been used to investigate the presence and ultrastructure of the mitochondria isolated by this procedure. The immunocytochemistry was carried out using cells containing wild-type levels of SSPI protein and cells over-expressing the protein. These results also demonstrate the capacity of mitochondria to import increased levels of protein in vivo. In vitro import experiments using COXIV-DHFR indicate that purified S. pombe mitochondria can efficiently import this precursor, and that protein translocation is dependent upon an oxidizable substrate and a membrane potential.
    Additional Material: 5 Ill.
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  • 132
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. 973-975 
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; genome mapping ; RNA helicase genes ; chromosome IV ; chromosome XI ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An ‘electroinic’ gene mapping procedure based on computer-aided search for overlapping gene sequences was used to identify adjacent genes and localize several putative RNA helicase genes to different chromosomes. PRP28 and AMD1 genes map to the right arm of chromosome IV netxt to sup2, which encodes a tyrsine tRNA PRP16, previouslymapped to chromosome XI, is tightly linked to MRP - L20. PRP22 is adjacent to PRE1, whose chromosomal location is currently unknown. The utility of this aproach in yeast gene mapping is evaluated.
    Additional Material: 1 Ill.
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  • 133
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XI ; CCEI ; shotgun sequencing ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the wequence of a 9·3 kb DNA segment of chromosome XI of Saccharomyces cerevisiae, located between the MAK11 locus and the centromere. This sequence contains four long open reading frames (ORFs). YKL160, YKL162, YKL164, YKL165 and part of another ORF, YKL166, covering altogether 90% of the entire sequence. One of these ORFs YKL164, corresponds to CCE1. Translation products of two other ORFs, YKL160 and YKL165, exhibit homology with previously known S. cerevisiae proetins: the robosomal protein L10, and the MYO2 gene product, respectively.
    Additional Material: 6 Ill.
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  • 134
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 8 (1992), S. 1025-1031 
    ISSN: 0749-503X
    Keywords: Saccharomycescerevisiae ; malic acid ; transport ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The transport system for malic acid present in Schizosaccharomyces pombe cells, growing in batch culture on several corbon sources, has been studied. It was found that the diarboxylic acid crrier of S. Pombe is a proton-dicarboxylate symporter that allows transport and accumulation as a function of ΔpH with the following kenetic parameters at pH 5·0: Vmax = 0·01 nmol of total malic acids -1 mg (dry weight) of cells, -1and Km = 0·1mM total malica acid uptake (pH 5·0) was accompanied by desappearance of extracellular protons, the uptake rates of which followed Michaelis-Menten kinetics as a function of the acid conscentration. The Km values, calculated as the concentrations either of anions or of undissociated acid, at various extracellular pH values, pointed to the monoanionic form as the transported species. Furthermore, accumulated free acid suffered rapid efflux after the addition of the portonophore carbonyl cyanid m-chlorophenyl hydrazone. These results suggested that the transport system was a dicarboxylateproton symporter. Growth of cells in a medium wiht glucose (up to 14%, w/v) and malic acid (1·5%, w/v) also resulted in proton-dicarboxylate activity, suggesting that the system, besides being constitutive, was still active at high glucose concentratons. The following dicarboxylic acids acted as competitive inhibitors of malic acid transport at pH 5·0: D- malic acid, succinic acid, fumaria acid oxaloacetic acid, α-Ketoglutaric acid, maleic acid, maleic and malonic acid. In addition all of these dicarboxylic acids induced proton movements that followed Michaelis-Menten kinetics. It was concluded that the malic negatively charged form (probably the monoanionic form) was transported by a proton-symport mechanism and that the carrier appeared to be a common ‘dicarboxylat transport sysmem’. The undissociated acid entered the cells slowly by simple diffusion.
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  • 135
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    Yeast 8 (1992), S. 1065-1075 
    ISSN: 0749-503X
    Keywords: Candida tropicalis ; auxotrophic mutants ; DNA transformation ; replicative and integrative plasmid ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The alkane-assimilating yeast Candida tropicalis was used as a host for DNA transformations. A stable ade2 mutant (Ha900) obtained by UV-mutagenesis was used as a recipient for different vectors carrying selectable markers. A first vector, pMK 16, that was developed for the transformation of C. albicans and caries an ADE2 gene marker and a Canadida autonomously replicating sequence (CARS) element promoting autonomous replication, was compatible for transforming Ha900. Two transformant types were observed: (i) pink transformants which easily lose pMK 16 under non-selective growth conditions; (ii) white transformants, in which the same plasmid exhibited a higher mitotic stability. In both cases pMK 16 could be rescued from these cells in Escherichia coli. A second vector, pADE2, containing the isolated C. tropicalis ADE2, gene was used to transform Ha900. This vector integrated in the yeast genome at homologous sites of the ade2 locus. Different integration types were observed at one or both ade2 alleles in single or in tandem repeats.
    Additional Material: 6 Ill.
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  • 136
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    Yeast 8 (1992) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 137
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    Yeast 8 (1992), S. S1 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 138
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    Yeast 8 (1992), S. S293 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 139
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    Yeast 8 (1992), S. S311 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 140
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    Yeast 8 (1992), S. S495 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 141
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    Yeast 8 (1992), S. S629 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 142
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    Yeast 8 (1992), S. 21-40 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 143
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    Yeast 8 (1992), S. 41-60 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 144
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    Yeast 8 (1992), S. 1-23 
    ISSN: 0749-503X
    Keywords: Saccharomyces ; taxonomy ; yeasts ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Additional Material: 10 Ill.
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  • 145
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; platelet derived endothelial cell growth factor ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A platelet-derived endothelial cell growth factor cDNA has been cloned, sequenced and expressed using the Saccharomyces cerevesiae PRBI promoter. Soluble recombinant platelet-derived endothelial cell growth factor constituted 0·5-1·0% of total soluble protein. Yeast soluble protein extracts containing recombinant platelet-derived endothelial cell growth factor stimulate the growth of calf palmonary artery endothelial cells in vitro.
    Additional Material: 2 Ill.
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  • 146
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    Yeast 8 (1992), S. 25-38 
    ISSN: 0749-503X
    Keywords: Yeasts ; flocculation ; FLO genes ; dsRNA ; Saccharomyces cerevisiae ; brewers' yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Yeast floccultion results from surface expression of specific proteins (lectins). Two flocculation phenotypes were suggested by physiological and biochemical tests, whereas genetic data suggested a larger number of mechanisms of flocculation. After reviewing the biochemistry, physiology and genetics of flocculation, a new hypothesis combining the data available from these different sources, is proposed.Flocculation results when lectins present on flocculent cell bind sugar residues of neighbouring cell walls. These sugar receptors are intrinsic to the mannan comprizing cell walls of Saccharomyces cerevisiae. Two lectin phenotypes were revealed by sugar inhibition studies. The gluco- and mannospecific NewFlo phenotype is not, as yet, found in genetically defined strains. Mannospecific flocculation (Flo 1 phenotype) is found in strains containing the genes FLO1, FLO5 and FLO8. This phenotype is also found following mutation of the TUP1 or CYC8 loci, in previously non-flocculent strains. It is therefore proposed that the structure gene for mannospecific flocculation is common or possibly unbiquitous in non-flocculent strains and in consequence, FLO1, FLO5 and FLO8 are probably regulatory genes, exerting positive control over the structure gene.Flocculation expression requires lectin secretion to the cell surface. Many of the observed ‘suppressions’ of flocculation may be due to mutations of the secretory process, involved in transporting structural proteins to the cell wall.The possible involvement of killer L double-stranded RNA with flocculation is suggested, given the lectin properties of viral coat proteins nad an association between L double-stranded RNA and the Flo 1 phenotype.
    Additional Material: 5 Ill.
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  • 147
    ISSN: 0749-503X
    Keywords: Yeast ; chromosome III ; CIT2 ; SUF2 ; tRNA Asn ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The complete nucleotide sequence of the D10H fragment (10850 bp) was determined. The D10H fragment is located on the right arm of chromosome III near the centromere and contains the SUF2 gene. Six open reading frames (ORFs) larger than 300 bp were found. One of them is the CIT2 gene encoding the cytoplasmic citrate synthase. The others are new putative genes and show no significant similarly with any known gene. In addition two tRNA genes (Asn and Pro) and a solo delta element were identified. Two ORFs were disrupted; no peculier phenotype was observed.
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  • 148
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    Yeast 8 (1992), S. 117-120 
    ISSN: 0749-503X
    Keywords: Thioredoxin ; TRX1 ; TRX2 ; genetic map location ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The two genes encoding thioredoxims in Saccharomyces cerevisiae, TRX1 and TRX2, map to chromosome XII and VII, respectively. From the DNA sequence of the intragenic region TRX1 is 500 bp downstream of PDC1. Tetrad analysis places TRX21·1 cM from ADE3, while a physical map of this region positions TRX2 4·5 kb downstreams of ADE3. The mapping of TRX1 adjacent to PDC1 clarifies previous results (Muller, E. G. D. J. Biol. Chem. 266, 9194-9202, 1991) that suggested a third thioredoxims gene.
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  • 149
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    Yeast 8 (1992), S. 95-106 
    ISSN: 0749-503X
    Keywords: Small heat-shock protein ; Hsp26 overexpression ; yeast (Saccharomyces cerevisiae) ; heat-shock proteins ; Hsp26-containing high molecular weight aggregate ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Hsp26 is one the major small heat-shock proteins (Hsp) of the Saccharomyces cerevisiae. yet its cellular role remains to be discovered. To examine the cellular consequences of overexpression of Hsp26, the gene encoding this protein (HSP26) was overexpressed from a multicopy plasmid using either its own promoter or by coupling it to the effecient constitutive PGK promoter. The PGK promoter provided the opportunity to overexpress Hsp26 under nonstress conditions and such high level synthesis, prior to a lethal heat shock (50°C), gave a small but reproducible elevation in thermotolerance. In transformed strains overexpressing Hsp26 under either stressed or non-stress conditions, the Hsp26 polypeptide was recovered almost exclusively as a high molecular weight aggregate. This high molecular weight aggregate (or heat-shock granule, HSG) was purified by differential centrifugation and sucrose gradient density centrifugation and shown, by electron microscopic analysis, to be of a uniform size (15-25 nm diameter). Analysis of the purified HSG demonstrated that it had a molecular weight of 550 kDa, yet contained no other integral polypeptides or other macromolecules.
    Additional Material: 7 Ill.
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  • 150
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome III ; sup61 ; RADI18 ; chromosome sequening ; Zn finger proteins ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A 7965 bp DNA segment from the right arm of chromosome III of Saccharomyces cerevisiae, encompassing the sup61 and RAD18 genes, was sequenced. Four new open reading frames were found in this DNA fragment. One of them YCR103, is 51% homologous with the G10 gene product of Xenopus laevis.
    Additional Material: 5 Ill.
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  • 151
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    Yeast 8 (1992), S. 171-182 
    ISSN: 0749-503X
    Keywords: Fission yeast ; pre-mRNA splicing ; intron architecture ; splice sites ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The architectural features of 73 introns found in 36 genes of the fission yeast Schizosaccharomyces pombe have been compiled and tabulated. The intron features of Saccharomyces cerevisia and other eukaryotes. The results that S. pombe displays quite different architectural features than the budding yeast S. cerevisiae. However, particularly in the 3′ region, S. pombe introns also appear to differ from mammalian introns.
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  • 152
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    Yeast 8 (1992), S. 223-225 
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome VI ; tRNA gene ; SUP11 ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Additional Material: 1 Ill.
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  • 153
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    Yeast 8 (1992) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 154
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; cell-cycle genes ; DNA synthesis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Two Saccharomyces cerevisiae genes previously unknown to be required for DNA synthesis have ben identified by screening a collection of temperature-sensitive mutants. The effects of mutations in DNA43 and DNA52 on the rate of S phase DNA synthesis were detected by monitoring DNA synthesis in synchronous populations that were obtained by isopycnic density centrifugation. dna43-1 and dna52-1 cells undergo cell-cycle arrest at the restrictive temperature (37°C), exhibiting a large-budded terminal phenotype; the nuclei of arrested cells are located at the neck of the bud have failed to undergo DNA replication. These phenotypes suggest that DNA43 and DNA52 are required for entry into or completion of S phase. DNA43 and DNA52 were cloned by their abilities to suppress the temperature-sensitive lethal phenotypes of dna43-1and dna52-1 cells, respectively. DNA sequence analysis suggested that DNA43 and DNA52 encode proteins of 59.6 and 80.6 kDa, respectively. Both DNA43 and DNA52 are essential for viability and genetic mapping experiments indicate that they represent previously unidentified genes: DNA43 is located on chromosome IX, 32 cM distal from his5 and DNA52 is located on chromosome IV, 0.9 cM from cdc34.
    Additional Material: 8 Ill.
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  • 155
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    Yeast 8 (1992), S. 39-45 
    ISSN: 0749-503X
    Keywords: Flow cytometry ; autolytic mutants ; protoplasts ; yeast ; viability assay ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Simple methods, based on the technique of flow cytometry, have been developed for the phenotypic characterization of yeast autolytic mutants and for the analysis of the formation and regeneration of the yeast protoplasts. The expression of lytic mutations determined uptake of the fluorescent dye propidium iodide, which could be carefully monitored by flow cytometry. Mixed populations of lysed and viable cells were precisely quantified and sorted, and the technique was also applied to demonstrate protection from lysis of mutant cells with cell wall defects, in the presence of osmotic stabilizers. Protoplast formation and regeneration was monitored by analysing relative cell size; this was facilitated by the preparation of homogeneous protoplast preparations. The technique of flow cytometry proved superior to other conventional methods for these types of study.
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  • 156
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    Yeast 8 (1992) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 157
    ISSN: 0749-503X
    Keywords: Maltose ; galactose ; raffinose fermentation ; nculeo-mitochondrial interaction ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The IMP2 gene of Saccharomyces cerevisiae is involved in the nucleo-mitochondrial control of maltose, galactose and raffinose utilization as shown by the inability of imp2 mutants of grow on these carbon sources in respiratory-deficient conditions or in the presence of ethidium bromide and erythromycin. The negative phenotype cannot be scored in the presence of inhibitors of respiration and oxidative phosphorylation, indicating that the role of the mitochondria in the utilization of the above-mentioned carbon sources in imp2 mutants is not at the energetical level.Mutantions in the IMP2 gene also confer many phenotypic alterations in respiratory-sufficient conditions, e.g. leaky phenotype on oxidizable carbon sources, sensitivity to heat shock and sporulation deficiency.The IMP2 gene has been cloned, sequenced and disrupted. The phenotype of null imp2 mutants is indistinguishable from that of the originally isolated mutant.
    Additional Material: 5 Ill.
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  • 158
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome II ; RAD16 ; DNA helicase ; double-finger motif ; DNA excision repair ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have analysed a region some 30 kb centromere distal form PHO5 on the right arm of yeast chromosome II and determined the nucleotide sequence of a 8.95 kb DNA segment from this region. By this analysis we were able to derive the precise location and the transcriptional orientation of CMD1, ALG1, SSN6 and LYS2. An open reading frame of 2370 bp was locatlized between SSN6 and LYS2, which has recently been identified (Schild et al., 1991) to be the RAD16 gene. The putative gene product, 790 amino acids in length, reveals several interesting freatures. It contains a nuclear target singnature and shares several blocks of similarity with the yeast recombinational repair protein RAD54 and the nuclear factor SNF2 (SW12), which is required for teh transcriptioal activation of a number of yeast genes. The similarity blocks in these three proteins are reminiscent of those found in the helicase superfamily. Furthrmore, RAD16 contains a novel ‘double-finger’ motif, which has been encountered in a variety of proteins from different organisms that are suggested to interact with DNA and are involved in diverse functions including site-specific recombination, DNA repair, and transcriptional regulation. The putative gene product of RAD16 then is the first example of a proteins in which the novel double-finger motif is found to be combined with a poteintial DNA helicase framework.
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  • 159
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    Yeast 8 (1992), S. 423-488 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Additional Material: 13 Ill.
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  • 160
    ISSN: 0749-503X
    Keywords: Protein glycosylation ; Saccharomyces cerevisiae ; outer chain mannosylation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have isolated two temperature-sensitive Saccharomyces cerevisiae mutants which exhibit a deficiency in mannose outer chain elongation of asparagine-linked oligosaccharide. The size of yeast glycoprotein, secretery form of invertase, of one mutant (ochl) was slightly larger than that of the sec18 mutant at the non-permissive temperature, while that of the other mutant (och2) was almost the same as that of the sec18 mutant. Unlike sec mutants, the och mutants were not deficient in secretion of invertase. The och1 mutant showed a 2+:2- cosegregation with regard to the temperature sensitivity and mannose outer chain deficiency, suggesting that a single gene designated as OCHI is responsible for these two phenotypes. The och1 mutant stopped its growth at the early stage of bud formation and rapidly lost its viability at the non-permissive temperature. The och1 mutation was mapped near the ole1 on the left arm of chromosome VII. The och1 mutant cells accumulated the external invertase containing a large amount of core-like oligosaccharides (Man9-10GlcNAc2) and a small amount of high mannose oligosaccharides (〉Man50GlcNAc2) at the non-permissive temperature. Production of the active form of human tissue-type plasminogen activator was increased in the och1 mutant compared with the parental strain, suggesting the potential advantage of this mutant for the production of mammalian-type glycoproteins which lack mannose outer chains in yeast.
    Additional Material: 8 Ill.
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  • 161
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: As part of the EEC project to sequence the entire chromosome III of Saccharomyces cerevisiae we have sequenced a total of 11,040 bp from near the right end of the chromosome. A new protein kinase gene was found at one extremity of the sequenced region (Wilson et al., 1992), while the previouslysequenced actin binding protein gene, ABPI, (Drubin et al., 1990) was found at the other extremity. We present here the sequence of the region between these two genes which has the potential to code for two new open reading frames (ORFs).
    Additional Material: 3 Ill.
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  • 162
    ISSN: 0749-503X
    Keywords: Mercaptoethanol ; dithiothreitol ; plasmalemma ; tonoplast ; H+-ATPase ; H+-permeability ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Mercaptoethanol and dithiothreitol (DTT) inhibited the acidification of external medium by by Saccharomyces Carlsbergensis cells and protoplasts during glucose oxidation. The inhibition was also observed when cells were incubated with mercaptoethanol or when mercaptoethanol and DTT were used to prepare protolasts. Experiments with S. carlsbergensis plasma membrene vesicles and vacuoles showed these thiol reagents to inhibitATP-dipendent generation of ΔpH and Em across plasma membrane vesicles and vacuoles but to activate their H+-ATPases. Mercaptoethanol and DTT are suggested to de-energize plasmalemma as well as tonoplast by increasing their H+-permeability and to disturb the cell ion homeostasis.
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  • 163
    ISSN: 0749-503X
    Keywords: DNA sequencing ; chromosome XI ; centromere CEN11 ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A 24·7 kb segment of the cosmid clone pUKG047 containing a Sau3AI-partial fragment from the centromere region of Saccharomyces cerevisiae chromosome XI was sequenced and analysed. A mexed strategy of directed methods including exonuclease III nested deletion, restriction fragment subcloning and oligonucleotide-directed sequences was carried out. Exclusive use waas made of the Applied Biosystems Taq DyeDeoxy™ Terminator Cycle technology and a laser-based ABI373A sequencing system for reactions, gel electrophoresis and automated reading. A total of 12 open reading frames (ORFs) was found. Nine new ORFs (YK102 to YK110) were identified, three of which (YK102, YK107, YK108)showed homologies to proteins of known function from other organisms. In addition, sequence analysis reveled three recently functionally characterized genes (MET14, VPS/SPO15, PAP1), which could be joined to the earlier published CEN11 region.
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  • 164
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    Yeast 8 (1992), S. 787-790 
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; ATPase ; chromosome V ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have sequenced a gene that encodes a 377 amino acid putative protein with an ATPase motif typical of the protein family including SEC18p (NSF = N-ethyl maleimide-sensitive fusion protein; vesicle-mediated endoplasmic reticulum to Golgi protein transfer), PAS1p (peroxisome assembly), CDC48p (VCP = valosin-containing protein; cell cycle) and TBP1 (Tat-binding protein). This gene, AFG1 for ATPase family, gene, also has substantial homology to these proteins outside the ATPase domain. AFG1 is located on chromosome V imediately centromere-proximal to M AK10.
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  • 165
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    Yeast 8 (1992), S. 791-799 
    ISSN: 0749-503X
    Keywords: Fission yeast ; Schizosaccharomyces pombe ; vacuolar ATPase ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The genes coding subunits A (vma1) and B (vma2) of the vacuolar H+-ATPase from Schizosaccharomyces pombe were cloned by hybridization to cDNAs of the homologous genes in Neurospora crasa. Both genes are interrupted by introns, two in vma1 and four in vma2. Positions of introns do not appear to be conserved when compared to those of N. crassa. The subunit A gene encodes a single product of 619 amino acids and is not interrupted by the coding sequence for a second product as found for Saccharomyces cerevisiae (Kane, P.K., Yamashiro, C.T., Wolczyk, D.F., Neff, N., Goebl, M., and Stevens, T.H. (1990). Science 250, 651-657).
    Additional Material: 4 Ill.
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  • 166
    ISSN: 0749-503X
    Keywords: Genome sequencing ; Saccharomyces cerevisiae ; chromosome II ; PDR1 ; multidrug resistance ; Zn binuclear cluster ; Leu zipper ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A 12,684 bp DNA fragment, between FUS3 and the centromere, from the left arm of chromosome II of Saccharomyces cerevisiae was sequenced as part of the European project to sequence the whole chromosome. This segment contains at least five complete new open reading frames (ORFs) and the beginning (191 first 5′ codons) of an ORF whose putative translational product is highly similar to the multidrug resistance PDR1 gene previously characterized by Balzi et al. (1987) on chromosome VII.
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  • 167
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 168
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    Yeast 8 (1992), S. 997-1006 
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; Permeases ; transport ; allantoin ; uracil ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have determined the structure of the allantoin permease (DAL4) gene of Saccharomyces cerevisiae. The gene patatively encodes a hydrophobic protein with a Mr of 71 755. It possesses the alternating hydrophobic-hydrophilic regions similar to those found in many other integral membrane proteins. The most striking feature of the allantoin permease component encoded by DAL4 is its striking similarity to the uracil permease component encoded by FUR4 Although data available indicate that these proteins do not share any overlap of function, their predicted protein sequences are 68% identical, 81% similar, and their DNA sequences are 70% identical. The upstream regulatory region of DAL4 contains all lof the characterized cis-acting elements previously reported for inducible allantoin pathway genes: six sequences homologous to UAS NTR, the element responsible for nitrogen catabolite repression-sensitive activation of allantoin pathway gene expression, and two sequences homologous to the cis-acting element responsible for inducere-responsiveness of the allantoin pathway genes, UIS. The finding of these homologous sequences predicted to exist on the basis of DAL4's expression characteristics, supports and strengthens the suggestion that these elements mediate the functions we have we have previously ascribed to them.
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  • 169
    ISSN: 0749-503X
    Keywords: Acetyl-coenzyme A synthetase ; carbohydrate metabolism ; Saccharomyces cerevisiae ; chromosome I ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A DNA fragment of Saccharomyces cerevisiae with high homology to the acetyl-coenzyme A (acetyl-CoA) synthetase genes of Aspergillus nidulans and Neurospora crassa has been cloned, sequenced and mapped to chromsome I. It contains an open reading frame of 2139 nucleotides, encoding a predicted gene product of 79·2 kDa. In contrast to its ascomycete homologs, there are no introns in the coding sequence. The first ATG codon of the open reading frame is in an unusual context for a translational start site, while the next ATG, 24 codons dowunstream, is in a more conventional context. Possible implications of two alternative traslational start sites for the celular lacalization of the enzyme are disussed. A stable mutant of this gene, obtained by the gene disruptiona technique, had the same low basal acitivity of acetyl-CoA synthetase as wild-type cells when grown on glucose but completely lacked the strong increase in activity upon entering the stationary phase, providing direct proof that the gene encodes an inducible acetlyl-CoA synthetase (ACSI) of yeast. As expected, the mutant was unable to grow on acetate as sole carbon source. Nevertheless, it showed normal induction os isocitrate lyase on acetate media, indicating that activity of acetyl-CoA sytheatase is dispensable for incuctionof the glyoxylate cycle in S. cerevisiae. Surprisingly, disruption of the ACSI gene did not affect growth on media containing ethanol as the sole crbon source. Demonstrating that threre are alternative pathways pathways leading to acetyl-CoA under these condintions.
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  • 170
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    Yeast 8 (1992), S. 1101-1103 
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; centromere V ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The WBP1 locus, encoding an essential component of the N-oligosaccharyl transferase, was mapped both genetically and physically. The gene is located on chromosome V between CENV and gcn4. The distance from CENV sequences is 2 kb.
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  • 171
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    Yeast 8 (1992), S. i 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 172
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    Yeast 8 (1992), S. S223 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 173
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    Yeast 8 (1992), S. S351 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 174
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    Yeast 8 (1992), S. S533 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 175
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    Yeast 8 (1992), S. S665 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 176
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome II ; ARS ; ACH1 ; FUS3 ; PEP1 ; vacuolar protein sorting ; carboxypeptidose Y ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the nucleotide sequence of an 11.4 kb DNA segment from the left arm of Saccharomyces cerevisiae chromosome II. This sequence contains a typical structure of a functional ARS as well as five open reading frames (ORFs) longer than 300 bp. One is PEP1, a gene encoding a transmembrane protein of 1579 amino acids which transits through the secretory pathway and is involved in vacuolar protein sorting. Two genes were previously sequenced: ACH1 (Lee et al., 1990) and FUS3 (Elion et al., 1990), which encode an acetyl-CoA hydrolase and a protein kinase involved in the cell division cycle, respectively. The last two ORFs localized on the complementary strand of ACH1 are not likely to be expressed.
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  • 177
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 178
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    Developmental Genetics 13 (1992), S. 111-117 
    ISSN: 0192-253X
    Keywords: Tetrahymena ; copy number ; histone H4 ; macronuclear DNA molecules ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In Tetrahymena, the DNA of the macronucleus exists as very large (100 to 4,000-kb) linear molecules that are randomly partitioned to the daughter cells during cell division. This genetic system leads directly to an assortment of alleles such that all loci become homozygous during vegetative growth. Apparently, there is a copy number control mechanism operative that adjusts the number of each macronuclear DNA molecule so that macronuclear DNA molecules (with their loci) are not lost and aneuploid death is a rare event. In comparing Southern analyses of the DNA from various species of Tetrahymena using histone H4 genes as a probe, we find different band intensities in many species. These differences in band intensities primarily reflect differences in the copy number of macronuclear DNA molecules. The variation in copy number of macronuclear DNA molecules in some species is greater than an order of magnitude. These observations are consistent with a developmental control mechanism that operates by increasing the macronuclear copy number of specific DNA molecules (and the genes located on these molecules) to provide the relatively high gene copy number required for highly expressed proteins. © 1992 Wiley-Liss, Inc.
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  • 179
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    Developmental Genetics 13 (1992), S. 126-132 
    ISSN: 0192-253X
    Keywords: Macronucleus ; macronuclear development ; Rf ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: During vegetative, asexual reproduction in heterozygous Tetrahymena thermophila, the macronucleus divides amitotically to produce clonal lineages that express either one or the other allele but not both. Because such phenotypic assortment has been described for every locus studied, its mechanism has important implications concerning the development and structure of the macronucleus. The primary tools to study assortment are Rf/ the rate at which subclones come to express a single allele stably, and the output ratio, the ratio of assortee classes. Because Rf is related to the number of assorting units, a constant Rf for all loci suggests that all genes are maintained at the same copy number. Output ratios reflect the input ratio of assorting units, with a 1:1 output ratio implying equal numbers of alleles at the end of macronuclear development. Because different outcomes would suggest a different macronuclear structure, it is crucial that these parameters be accurately measured. Although published Rf values are similar for all loci measured, there has been no commonly accepted form of presentation and analysis. Here we examine the experimental determination of Rf. First, we use computer simulation to describe how the variability inherent in the assortment process affects experimental determination of Rf. Second, we describe a simple method of plotting assortment data that permits the uniform calculation of Rf, and we describe how to measure Rf accurately in instances when it is possible to score only the recessive allele. Using this method to produce truly comparable Rfs for all published data, we find that most, if not all, loci assort at Rfs consistent with ∼45 assorting units, as has been asserted. © 1992 Wiley-Liss, Inc.
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  • 180
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    Developmental Genetics 13 (1992), S. 118-125 
    ISSN: 0192-253X
    Keywords: Microinjection ; macronucleoplasm ; transformation ; Paramecium ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Transformation by microinjection of macronucleoplasm in Paramecium caudatum was investigated. Macronucleoplasm with three genetic markers (behavior, trichocyst, and mating type) was injected into the macronucleus. To facilitate microinjection, in most cases, paramecia were immobilized in a gelatin (7.5%) solution. The injected cells began to express a dominant gene (cnrA+ or cnrB+) of the donor 9-24 hr after injection. Expression did not require cell division suggesting injected macronucleoplasm was capable of expressing a phenotype. The amount of injected macronucleoplasm appears to correlate with the frequency of successful expression but not to correlate with the time required for expression. After a number of fissions, the injected cells produced clones which had cells expressing the phenotype of the donor. This suggests that injected macronucleoplasm was replicated and expressed in the recipient cell lines. The transformed clones were classified into two groups. In one group, transformation was stable. All cell lines derived from the injected cells expressed a phenotype similar to the heterozygote of donor and recipient cells. In the other group, transformation was unstable. During the first five to seven fissions after injection, at each division, cells produced one daughter cell which later reverted to the recipient phenotype. After this unstable period, cells no longer produced the recipient phenotype but produced the donor phenotype exclusively. Donor and recipient phenotypes were, thus, segregated in different cell lines. Observation of genetic markers and analysis by computer simulation shed light on the mode of transmission of injected macronucleoplasm. In stable transformation, injected macronucleoplasm appears to be distributed equally to daughter cells. In unstable transformation, injected macronucleoplasm is distributed only to one of the daughter cells at every division until about the fifth to seventh fission after injection and then begins to assort equally to daughter cells. The cell cycle stage at injection may influence the mode of transformation. Interspecific microinjection of macronucleoplasm from P. multimicronucleatum and P. tetraurelia to P. caudatum. resulted in the expression of foreign genes in P. caudatum. In one case, injection of macronucleoplasm of P. tetraurelia produced a stable transformant indicating replication of foreign macronucleoplasm in P. caudatum. This work reveals the mode of transformation by injected macronucleoplasm and shows the possibility of transformation among Paramecium species, which is significant in the study of the conservation of gene products and the mechanism of gene expression in different species. © 1992 Wiley-Liss, Inc.
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  • 181
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    Developmental Genetics 13 (1992), S. 143-150 
    ISSN: 0192-253X
    Keywords: Extrachromosomal nucleoli ; in vitro transcription ; chromatin ; ribosomal DNA (rDNA) ; Tetrahymena ; P1 nuclease mapping ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An in vitro transcription system was established using extrachromosomal nucleoli from Tetrahymena pyriformis macronuclei as a template. Ribosomal precursor RNA (pre-rRNA) and nascent pre-rRNA chains were removed from isolated nucleoli by treatment with RNase T1 and Sarkosyl. Nucleoli were then incubated in a RNA synthetic cocktail containing a cellular extract from Tetrahymena thermophila. The transcription product was examined for the presence of transcripts from T. pyriformis ribosomal DNA (rDNA) by P1 nuclease protection mapping using a DNA probe from a T. pyriformis rDNA clone. A sequence difference between T. pyriformis and T. thermophila in the 5′ region of their 35S pre-rRNAs permitted exclusive detection of T. pyriformis transcripts. The results showed that faithful transcription initiation occurred in vitro from the in vivo initiation site of the nucleolar template and that the nucleolar template had a much higher efficiency of transcription than that of the purified rDNA clone. This system may offer unique advantages for future studies of tran-scriptional control during development and differentiation. © 1992 Wiley-Liss, Inc.
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  • 182
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    Developmental Genetics 13 (1992), S. 160-166 
    ISSN: 0192-253X
    Keywords: Deletion mapping ; exocytosis ; genetic complementation ; genomic exclusion ; mucocyst ; nullisomic strains ; regulated secretion ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Under appropriate conditions, Alcian Blue-induced exocytosis of Tetrahymena mucocysts leads to formation of a capsule that surrounds the cell. This phenomenon is an example of regulated secretion, a mechanism of fundamental significance in eukaryotic cells. In order to dissect genetically the mechanism of mucocyst biogenesis and regulated exocytosis, mutants unable to form capsules (Caps-) were isolated. In this paper we report a genetic characterization of Caps- mutants in this collection. The mutations in mutants SB255 and SB281 behave as single recessive Men-delian mutations. The mutation in SB251 is restricted to the macronucleus, and could not be further characterized by the genetic methods we used. Complementation tests suggest the existence of at least 2 genes, named exoA and exoB; additional mutant loci are likely to be included in the mutant collection. Deletion mapping using nulli-somic strains showed that exoA and exoB are located on the left arm of chromosome 4. The exo-3 mutation, which behaves as recessive and complements with exoA1 in SB255 and exoB2 in SB281, maps to chromosome 3. These Caps- mutants may be useful for the elucidation of the developmental pathway of mucocyst biogenesis and the control of regulated secretion in eukaryotic cells. © 1992 Wiley-Liss, Inc.
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  • 183
    ISSN: 0192-253X
    Keywords: Acid hydrolases ; sec allele ; phenotype ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The genetics and phenotypic features (light and electron microscopy) of a secretory mutant, MS-1, of Tetrahymena thermophila blocked in secretion of lysosomal acid hydrolases have been analyzed. Although blocked constitu-tively in secretion, MS-1 contains active lysosomal hydrolases in amounts equivalent to the wild type. The 3:1 segregation in F-2 in sib crosses and the 1:1 segregation in test crosses indicate that the block in secretion of lysosomal hydrolases is controlled by a recessive single gene locus termed sec. The sec allele of MS-1 proved also to be responsible for the highly vacuolarized phenotype the mutant developed when it was transferred from nutrient medium into buffers of low ionic strength. Deletion mapping by crossing MS-1 with nullisomic strains, all secreting lysosomal hydrolases at wild-type rates, was performed. The sec phenotype was expressed in monosomic-4 progeny only, indicating that the sec allele is located on chromosome 4 of T. thermophila. © 1992 Wiley-Liss, Inc.
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  • 184
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    Developmental Genetics 13 (1992) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 185
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    Developmental Genetics 13 (1992), S. 181-186 
    ISSN: 0192-253X
    Keywords: Inheritance ; non-genic ; pattern-formation ; ciliate ; Stentor coeruleus ; Tartar ; Vance ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The one form of cytoplasmic inheritance that has not been assimilated into the Central Dogma is the inheritance of surface structural patterns, a phenomenon most clearly expressed in cilates. Vance Tartar, although he worked with a genetically undomesticated organism (Stentor coeruleus), provided early evidence for the crucial role of clonally propagated features of the cell cortex. He showed that the capacity for development of cortical organelle systems is associated with a particular relational feature, the “locus of stripe contrast” (LSC), and that clonally inherited cortical variants (homopolar doublets) could be created at will by microsurgical operations that duplicated the LSC. Tartar also hoped to demonstrate the existence of what David Nanney called “cellular architects” by provoking stentors to carry out entirely novel types of morphogenetic performances. He eventually acknowledged failure, although the bizarre juxtapositions by which he attempted to elicit such novel performances did bring about specific and illuminating defects in cortical development. Subsequent analyses of similar defects in other ciliates revealed not the unitary “pattern factor” postulated by Tartar, but rather a hierarchy of distinct patterning mechanisms. Nonetheless, by pursuing an embryological approach toward morphogenesis in a highly regulative ciliate, Tartar uncovered relational aspects of pattern-determination; this, in my view, delineates the major problem that we must solve to gain understanding of intracellular patterning. © 1992 Wiley-Liss, Inc.
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  • 186
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    Developmental Genetics 13 (1992), S. 203-209 
    ISSN: 0192-253X
    Keywords: Ciliate ; basal body ; Stentor ; size dependent regulation ; ultrastructure ; adoral zone of membranelles ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The number and length of oral membranelles were determined for both large and small Stentor from well-fed, growing cultures and nutrient-deprived cultures, respectively. Small cells possess both significantly fewer and shorter membranelles than do large cells. For both large and small cells, each membranelle is composed of three rows of basal bodies. The membranelles closest to the gullet have a third row that is only slightly shorter than the other two. The third row becomes rapidly shorter as membranelles become increasingly distant from the gullet. A short distance from the gullet, and for the remainder of the band, the third row is composed ofonly one to four basal bodies. The first two rows consist of approximately 35 basal bodies each in large cells and approximately 26 basal bodies each in small cells. This indicates that Stentor regulates the number of basal bodies per row, but not the number of rows, in response to changes in cell size. © 1992 Wiley-Liss, Inc.
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  • 187
    ISSN: 0192-253X
    Keywords: Ciliate ; cortex ; basal body ; morphogenesis ; pattern formation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The present study reveals a deficiency in the number of ciliated basal bodies along 180° rotated ciliary rows (IRs) in Tetrahymena. This feature is common to IRs recently generated in young clones with stable corticotypes (total number of ciliary rows per cell), irrespective of the number of IRs present per cell or their cellular location, and is found before the cell loses any of the IRs. In cells bearing three IRs, the IRs on the two sides of the inversion immediately next to normal ciliary rows (junctures) exhibit an even greater deficiency in ciliated basal bodies, compared to the IR located internally between two other IRs; the normal ciliary rows flanking the inversion are also somewhat deficient. These observations show that the IRs of Tetrahymena are structurally deficient, hence developmentally defective, and suggest that they are intrinsically unstable. We propose that basal body development along IRs tends to be truncated before the stage of ciliation; such basal bodies would fail to acquire the potential to serve as nucleating centers for new basal body development in the next round of basal body proliferation, leading to the eventual loss of the IRs. © 1992 Wiley-Liss, Inc.
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  • 188
    ISSN: 0192-253X
    Keywords: Cell division ; microinjection ; division plane ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Understanding the mechanism that determines the cell division plane is one of the most important problems in the fields of cell and developmental biology. Studying the timing and site of formation of contractile ring (CR) micro-filaments provides key information for solving the problem. We tried to create a nonfunctional CR in Tetrahymena by microinjecting rabbit skeletal muscle actin, which can copolymerize with Tetrahymena actin but has properties different from those of Tetrahymena actin. When skeletal muscle actin was injected in a predivision stage, before the onset of furrow constriction, long-term arrest of cell division was observed. Muscle actin did not cause any delay in cell division when the actin was injected at any stage other than the predivision stage. In all cases, muscle actin had little affect on other actin-related functions. Injected skeletal muscle actin polymerized near the equatorial division plane in cases of cell division arrest; it polymerized at other nonspecific locations when cell division was observed. Arrest occurred when the microinjection took place in the 17-min period just before the start of furrowing. This period coincides with the occurrence of equatorial deposits of p85, which is also suggested to be required for the determination of the division plane. The present experimental results are consistent with the idea that p85 is a crucial factor for determining the cell division plane and also functions as a polymerization nucleus for CR microfilaments. © 1992 Wiley-Liss, Inc.
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  • 189
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    Developmental Genetics 13 (1992), S. 229-234 
    ISSN: 0192-253X
    Keywords: Oral morphogenesis ; cc1 mutation ; cell cycle ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: This study examines the timing of micronuclear mitosis during the vegetative cell cycle and shows that mitosis begins early in the division process and coincides approximately with the earliest stages of oral morphogenesis (about 0.6 in the cell cycle in synchronous cell samples). The cc1 mutation blocks cell cycle progression prior to the point of commitment to division. Although the cc1 mutation blocks macronuclear DNA synthesis under restrictive conditions, it does not block micronuclear DNA synthesis. However, absence of functional cc1 gene product leads to blockage of micronuclear mitosis prior to completion of anaphase. This point coincides with commitment to division and is also the point at which oral morphogenesis is blocked in cc1 cells. The tim-ings of the transition points for micronuclear mitosis and oral morphogenesis in cc1 cells are closely associated in both synchronous cell samples and in asynchronous cultures. © 1992 Wiley-Liss, Inc.
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  • 190
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    Developmental Genetics 13 (1992) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 191
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    Developmental Genetics 13 (1992), S. 255-255 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 192
    ISSN: 0192-253X
    Keywords: Surface antigen ; Paramecium primaurelia ; macronuclear DNA ; DNA rearrangement ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The Paramecium primaurelia cell surface is covered with a high molecular weight protein called the surface antigen. Several genes encode alternative surface antigens, but only one is expressed at a time. In addition, each of these genes shows a high degree of allelic polymorphism. Paramecium primaurelia strains 156 and 168 have different alleles of the G antigen gene whose respective antigens can be distinguished in vivo using specific antibodies. An interallelic exclusion phenomenon has been previously described: 94% of the 156/168 heterozygotes express only the 156 allele of the G gene; 6% express both the 156 and the 168 alleles. The phenotype of the heterozygotes is determined at the time of macronuclear differentiation. We have investigated the molecular basis for the different heterozygous phenotypes. Both mRNAs are always produced, and the 156 mRNA is always more abundant than the 168 mRNA. The relative amounts of these messages, however, vary greatly between different heterozygotes and parallel their phenotype. Pushing the analysis further, we show that the copy number of each allele in the macronucleus correlates with the relative amounts of the mRNAs. However, allelic dosage alone is not sufficient to explain the variations of the mRNA ratio. The G antigen gene is located near a telomere in the macronucleus. We show that the distance between the 156G gene and the telomere is different in homozygotes and heterozygotes. It also varies among heterozygotes and is correlated with the mRNA ratio. Thus, we have identified two different parameters, both linked to the genome rearrangements occurring during macronuclear differentiation, that correlate with the relative expression of the two alleles. Two hypotheses concerning the influence of the telomere position on the expression of the gene are discussed. © 1992 Wiley-Liss, Inc.
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  • 193
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    Developmental Genetics 13 (1992), S. 326-330 
    ISSN: 0192-253X
    Keywords: Homeotic ; segmentation ; Pc-group genes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: polyhomeotic is a member of a group of genes, the Pc-group responsible for the maintenance of gene expression during development. In particular, the Pc-group of genes is involved in the correct expression of homeotic genes of the bithorax and Antennapedia complexes.Molecular analysis reveals that the Pc-group genes function relatively late in development, once homeotic gene expression has been correctly initiated. This initiation of homeotic gene expression depends on interaction between genes in the segmentation gene hierarchy, the gap and pair-rule genes.The in situ analysis presented here indicates that polyhomeotic transcripts are expressed in a spatially restricted fashion early in development. This blastoderm expression is under the control of genes in the segmentation hierarchy. Given these results, and the role of polyhomeotic in the correct maintenance of homeotic gene expression, I propose that polyhome otic expression may relay the initiation of homeotic gene expression with other mechanisms involved in the maintenance of this expression, involving the other Pc-group genes. © 1992 Wiley-Liss, Inc.
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  • 194
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    Developmental Genetics 13 (1992), S. 367-375 
    ISSN: 0192-253X
    Keywords: FLP-recombinase ; FRT ; embryonic mosaics ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An efficient method for generating embryonic mosaics using a yeast site-specific recombinase (FLP), under the control of a heat shock promoter, is described. FLP-recombinase can promote mitotic exchange between homologous chromosomes that contain FRT (FLP Recombination Target) sequences. To demonstrate the efficiency of FLP-recombinase to generate embryonic mosaics, clones of the recessive and cell autonomous mutation armadillo (arm), detected by their ability to differentiate ectopic denticles in the naked cuticle of each abdominal segment, have been induced. We have analyzed the parameters of FLP-recombinase induced embryonic mitotic recombination and have demonstrated that clones can be efficiently induced during the postblastoderm mitotic divisions. We discuss applications of this technique for the analyses of the roles of various mutations during embryonic patterning. © 1992 Wiley-Liss, Inc.
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  • 195
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 13 (1992), S. 359-366 
    ISSN: 0192-253X
    Keywords: Glutamine synthetase I ; femalesterile mutations ; D. melanogaster ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A female-sterile mutation (fs(2) PM11-19) was recovered in a screen for P-M hybrid dysgenesis induced mutations uncovered by a deletion of region 21B and was identified as an allele of the gene encoding the Drosophila glutamine synthetase I (GSI) mitochondrial isozyme.Molecular analysis has shown that fs(2)PM11-19 contains a 5 kb insert within 500 bp upstream of the transcriptional start site of the gsI gene. Mutant flies have extremely low levels of gsl transcription and GSI activity. A pre-existing deficiency (Df(2L) netpm1) with a breakpoint near the transcription start site was also found to be a female-sterile allele of gsl.All eggs laid by PM11-19 homozygous females, as well as by females heterozygous for this mutation and a deletion or any of several recessive lethal alleles of the gsl gene, fail to hatch. We conclude that an adequate level of maternally supplied GSI activity is necessary in the early stages of Drosophila embryonic development. © 1992 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
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  • 196
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 13 (1992) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 197
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 13 (1992), S. 376-386 
    ISSN: 0192-253X
    Keywords: Bronze-1 ; transposable element ; germinal reversion ; somatic excision ; microsporogenesis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Three characteristics of standard Mutator lines reflect developmental regulation: new mutants usually involve single gametes, somatic excision is restricted to terminal cell divisions during tissue development, and germinal excision is rare. By selection for earlier (larger) somatic sectors in the aleurone, a Mutator line was identified that exhibits a dramatic elevation in somatic excision frequency during the first three nuclear divisions of the endosperm and more than a 10-fold increase in germinal reversion from the bzl::Mul reporter gene. The programming of early sectoring is dominant in crosses with Mutator lines containing diverse reporter alleles. Germinal reversion is biased 5- to 10-fold for events through the pollen compared to the ear. The timing of germinal excision in the tassel is late because somatic excision sectors in the anthers are small; however, 98% of the germinal revertants are concordant. These observations indicate that in the early sectoring line Mu excision usually occurs before the mitotic divisions that separate gametic nuclei and may be restricted to the early stages of microsporogenesis. © 1992 Wiley-Liss, Inc.
    Additional Material: 2 Ill.
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  • 198
    ISSN: 0192-253X
    Keywords: Chromosome spreads ; meiotic mutants ; synaptonemal complex ; three-dimensional reconstruction ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have utilized spreading methods as well as serial sectioning three-dimensional reconstruction to examine meiotic chromosome behavior in cells homozygous for the rad3-1 mutation in Coprinus cinereus. Comparison of 42 wild-type nuclei that had been spread, stained with silver, and viewed by electron microscopy with 30 mutant nuclei treated in the same manner revealed several defects in the mutant. Axial core formation was defective in the mutant, although limited side-by-side association of axial cores was observed. To detect any differences in three-dimensional architecture between the wild-type and mutant nuclei, we reconstructed three of the former and six of the latter after serial sectioning. It was not possible to trace the expected number of axial cores from section to section in the mutant, although some tripartite synaptonemal complex was observed. Many axial core ends failed to terminate in the nuclear envelope in the mutant. This spectrum of defects (incomplete axial core assembly with some tripartite synaptonemal complex formation) had not been observed previously in either C. cinereus or other systems. We conclude that this combination of spreading and sectioning methods is very useful for analysis of meiotic mutants. © 1993 Wiley-Liss, Inc.
    Additional Material: 3 Ill.
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  • 199
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 13 (1992), S. 411-424 
    ISSN: 0192-253X
    Keywords: Maize ; mutations ; female ; meiosis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A modified enzyme digestion technique of ovary isolation followed by staining and squash preparation has allowed us to observe female meiosis in normal maize meiotically dividing megaspore mother cells (MMCs). The first meiotic division in megasporogenesis of maize is not distinguishable from that in mi-crosporogenesis. The second female meiotic division is characterized as follows: (1) the two products of the first meiotic division do not simultaneously enter into the second meiotic division; as a rule, the chalazal-most cell enters division earlier than the micropylar one, (2) often the second of the two products does not proceed with meiosis, but degenerates, and (3) only a single haploid meiotic product of the tetrad remains alive, and this cell proceeds with three rounds of mitoses without any intervening cell wall formation to produce the eight-nucleate embryo sac. This technique has allowed us to study the effects of five meiotic mutations (aml, aml-pral, afdl, dsy *-9101, and dvl) on female meiosis in maize. The effects of the two alleles of the aml gene (aml and aml-pral) and of the afdl and dsy *-9101mutations are the same in both male and female meiosis. The aml allele prevents the entrance of MMCs into meiosis and meiosis is replaced by mitosis; the aml-pral permits MMCs to enter into meiosis, but their progress is stopped at early prophase I stages. The afdl gene is responsible for substitution of the first meiotic (reductional) division by an equational division including the segregation of sister chromatid centromeres at anaphase I. The dsy * -9101 gene exhibits abnormal chromosome pairing; paired homologous chromosomes are visible at pachytene, but only univalents are observed at diakinesis and metaphase I stages. These mutation specific patterns of abnormal meiosis are responsible for the bisexual sterility of these meiotic mutants.The abnormal divergent shape of the spindle apparatus and the resulting abnormal segregation of homologous chromosomes observed in micro-sporogenesis in plants homozygous for the dv1 mutation have not been found in meiosis of megasporogenesis. Only male sterility is induced by the dv1 gene in the homozygous condition. © 1993 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
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  • 200
    ISSN: 0192-253X
    Keywords: Meiotic pairing ; rDNA ; Drosophila ; disjunction ; sex chromosomes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The Drosophila melanogaster ribosomal DNA (rDNA) functions as an X-Y meiotic pairing site. Deletions encompassing the X chromosomal rDNA block (located in the heterochromatin) disrupt X-Y pairing and disjunction. Insertions of single, complete rRNA genes at ectopic locations on the heterochromatically deficient X partially restore X-Y pairing capacity. This study was undertaken to test fragments of an rDNA repeat for the ability to stimulate X-Y pairing and disjunction and to test for relationships between pairing capacity and two other phenotypes associated with rDNA insertions: transcription and the ability to organize a nucleolus. Insertions of three different fragments, all of which retained the rDNA promoter and upstream spacer sequences and which differed among each other in the length of downstream sequences, were obtained by P-element mediated transformation. One of the fragments is truncated only 140bp downstream from the promoter. Insertions of all three fragments proved capable of stimulating X-Y disjunction. Double insertions were substantially more effective than single insertions. RNA/PCR analysis was used to show that transcripts initiated at the inserted rDNA promoters are present in testis RNA from all insertions. Treatment with an antinucleolar antibody revealed that none of the insertions was associated with a mininucleolus. Thus promoter-containing rDNA fragments are autonomously capable of being transcribed and of functioning as X-Y pairing sites, but not of forming a mini-nucleolus. © 1993 Wiley-Liss, Inc.
    Additional Material: 8 Ill.
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