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  • 1
    Electronic Resource
    Electronic Resource
    Determination of a specific region of the purine–cytosine permease involved in the recognition of its substrates (1992)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 6 (1992), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Three u.v.-induced mutants of the purine–cytosine permease gene of Saccharomyces cerevisiae, with altered apparent Michaelis constant of transport (Kmapp), were cloned and sequenced. One of the mutants had extensive nucleotide replacement, whereas the other two had a single mutation. To evaluate the contribution of the different amino acid replacements to the phenotype of the complex mutant, simpler mutants were created by site-directed mutagenesis. All the amino acid replacements found in the segment from amino acids 371 to 377 inclusive, contribute to the determination of the phenotype. According to the model postulated this segment lies on the cell surface. In particular, amino acids at position 374 and 377 modulate the affinity of the permease towards its substrates. In the wild-type, when asparagine is present at both of these positions, the lowest Kmapp values are found.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-2958.1992.tb01757.x
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  • 2
    Electronic Resource
    Electronic Resource
    Correlation between restriction map, genetic map and catalytic functions in the gene complex URA2 (1987)
    Springer
    Molecular genetics and genomics 209 (1987), S. 283-289 
    Details
    ISSN: 1617-4623
    Keywords: Deletion mapping ; Feedback inhibition ; Channelling ; Carbamylphosphate synthetase ; Aspartate transcarbamylase complex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We replaced the URA2 gene by six different deleted alleles constructed in vitro by Bg/II digestion in order to correlate the genetic map with the restriction map and to define the regions coding for the different functions of the carbamylphosphate synthetase — aspartate transcarbamylase complex (CPSase-ATCase). We also enlarged the collection of ura2 point mutations by using a positive selection method based on resistance to the toxic accumulation of ureidosuccinic acid (USA). Of the new independent mutations nine mapped in the intermediary zone, a previously defined mutationless region localized between regions coding for CPSase and ATCase. This shows that the former definition resulted from analysis of a limited number of mutants (40). The study of an allele deleted in the intermediary zone shows that this sequence codes for a protein region necessary for the feedback inhibition of the CPSase-ATcase enzyme complex. The CPSase- ATCase- phenotype of 26 mutants resistant to USA accumulation shows the importance of the in vivo channelling of carbamylphosphate in the CPSase-ATCase complex for USA and subsequent pyrimidine biosynthesis. Finally, our results confirm that the CPSase and ATCase activities are separate functions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00329655
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  • 3
    Electronic Resource
    Electronic Resource
    Reactivation of the ATCase domain of the URA2 gene complex: a positive selection method for Ty insertions and chromosomal rearrangements in Saccharomyces cerevisiae (1995)
    Springer
    Molecular genetics and genomics 246 (1995), S. 767-773 
    Details
    ISSN: 1617-4623
    Keywords: Ty retrotransposon ; Duplication ; Saccharomyces cerevisiae ; ATCase ; URA2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetic rearrangements such as deletions or duplications of DNA sequences are rarely detected in the yeast Saccharomyces cerevisiae. We have developed a screening system using the URA2 gene coding for the bi-functional CPSase-ATCase (carbamyl phosphate synthetase — aspartate transcarbamylase) to select positively for these kinds of events. Nonsense mutations in the CPSase region cause a complete loss of the ATCase activity because of their strong polar effect. Thirty-seven ATCase+ revertants were isolated from a strain containing three nonsense mutations in the proximal CPSase region. Genetic and structural analysis of the URA2 locus in these strains allowed us to characterize two major classes of revertants. In the first, an entire copy of a Ty transposon was found to be inserted in the CPSase coding domain. This event, which represents a new form of Ty-mediated gene activation was further analysed by mapping the Ty integration site in 26 strains. In a second class of revertants, we observed chromosomal rearrangements and, in particular, duplication of the ATCase region and its integration in a new chromosomal environment in which this sequence becomes active.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00290725
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  • 4
    Electronic Resource
    Electronic Resource
    The Characterization of Two New Clusters of Duplicated Genes Suggests a ‘Lego’ Organization of the Yeast Saccharomyces cerevisiae Chromosomes (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 861-869 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; cluster ; duplication ; genome sequencing ; evolution ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The systematic sequencing of 42 485 bp of yeast chromosome VII (nucleotides 377948 to 420432) has revealed the presence of 27 putative open reading frames (ORFs) coding for proteins of at least 100 amino acids. The degree of redundancy observed is elevated since five of the 27 ORFs are duplications of a previously identified gene. These duplicated copies may be classified in two types of cluster organization. The first type includes genes sharing a significant level of identity in the amino acid sequences of their predicted protein product. They are recovered on two different chromosomes, transcribed in the same orientation and the distance between them is conserved. The second type of cluster is based on one gene unit tandemly repeated. This duplication is itself repeated elsewhere in the genome. The level of nucleic acid identity is high within the coding sequence and the non-coding region between the two repeats. In addition, the basic gene unit is recovered many times in the genome and is a component of a multigene family of unknown function. These organizations in clusters of genes suggest a ‘Lego organization’ of the yeast chromosomes, as recently proposed for the genome of plants (Moore, 1995). The sequence is deposited in the Yeast Genome Databank under Accession Number from Z72562 to Z72586. © 1997 John Wiley & Sons, Ltd.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Sequence of a 9·8 kb segment of yeast chromosome II including the three genes of the MAL3 locus and three unidentified open reading frames (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 11 (1995), S. 667-672 
    Details
    ISSN: 0749-503X
    Keywords: chromosome II ; S288C ; MAL3 ; MAL1 ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the DNA sequence of a segment located on the right arm of chromosome II from Saccharomyces cerevisiae S288C near the subtelomeric sequences. The sequence was determined using a random cloning strategy followed by an oligonucleotide-directed sequencing. The segment contains four non-overlapping open reading frames (ORFs) YBR297w, YBR298c, YBR299w and YBR301c, and two overlapping ones (YBR300c and YBR300w). Three of them - YBR297w, YBR298c and YBR299w - are the MAL3R (transcriptional regulatory protein), MAL3T (maltose permease) and MAL3S (maltase) genes of the MAL3 locus previously localized. The three other ORFs are unidentified. Another MAL locus (MAL1) has been localized on chromosome VII. The Mal- phenotype of strain S288c cannot be explained by telomeric silencing. The sequences have been submitted to the EMBL data library under Accession Numbers Z36166; Z36167; Z36168; Z36169 and Z36171.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320110707
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  • 6
    Electronic Resource
    Electronic Resource
    Recovery of gene function by gene duplication in Saccharomyces cerevisiae (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 11 (1995), S. 169-177 
    Details
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome ; ATCase ; URA2 ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A prototroph revertant (Rev9) selected from an ATCase- mutant of the URA2 gene containing three nonsense mutations was shown to contain two ATCase coding sequences. We cloned both ATCase coding areas to show that the duplicated locus (dl9) was the only functional one. Its size corresponded roughly to the second half of the URA2 wild-type gene. Sequence analysis of the 5′ end of dl9 indicated that this duplicated sequence was inserted within the intergenic region close to the MRS3 gene and was transcribed from an unknown promoter divergently from the MRS3 gene. The event leading to the revertant strain Rev9 included a rearrangement that increased the size of chromosome X by about 60 kb. In agreement with such a rearrangement, recombination was undetectable in the vicinity of the locus dl9. Genetic mapping confirms that the MRS3 gene is 2 cM distal to the URA2 gene on the right arm of chromosome X.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320110208
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  • 7
    Electronic Resource
    Electronic Resource
    Analysis of 21·7 kb DNA Sequence from the Left Arm of Chromosome VII Reveals 11 Open Reading Frames: Two Correspond to New Genes (1997)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 13 (1997), S. 475-477 
    Details
    ISSN: 0749-503X
    Keywords: genome sequencing ; chromosome VII ; Saccharomyces cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The DNA sequence of a fragment of 21 731 bp (nucleotides 87408 to 109138) located on the left arm of chromosome VII from Saccharomyces cerevisiae S288C has been determined using a random cloning strategy followed by an oligonucleotide-directed sequencing. This fragment contains eight complete genes previously sequenced (CLG1, SKI8, VAM7, YPT32, MIG2, SIP2, SPT16 and CHC1), the 5′ part of POX1 and two other complete unidentified open reading frames of more than 100 amino acids. © 1997 John Wiley & Sons, Ltd.
    Additional Material: 1 Tab.
    Type of Medium: Electronic Resource
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