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  • 1
    Electronic Resource
    Electronic Resource
    In vitro mutagenesis of the mitochondrial leucyl-tRNA synthetase of S. cerevisiae reveals residues critical for its in vivo activities (1992)
    Springer
    Current genetics 22 (1992), S. 69-74 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondria ; Aminoacyl-tRNA synthetase ; RNA splicing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The mitochondrial leucyl-tRNA synthetase (mLRS) of Saccharomyces cerevisiae is involved in both mitochondrial protein synthesis and pre-mRNA splicing. We have created mutations in the regions HIGH, GWD and KMSKS, which are involved in ATP-, amino acid-and tRNA-binding respectively, and which have been conserved in the evolution of group I tRNA synthetases. The mutants GRD and NMSKS have no discernible phenotype. The mutants AWD and ARD act as null alleles and lead to the production of 100% cytoplasmic petites. The mutants HIGN, NIGH and KMSNS are unable to grown on glycerol even in the presence of an intronless mitochondrial genome and accumulate petites to a greater extent than the wild-type but less than 40%. Experiments with an imported bI4 maturase indicate that the lesion in these mutations primarily affects the synthetase and not the splicing functions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00351744
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  • 2
    Electronic Resource
    Electronic Resource
    A comprehensive compilation of 1001 nucleotide sequences coding for proteins from the yeast Saccharomyces cerevisiae (=ListA2) (1993)
    Springer
    Current genetics 23 (1993), S. 66-91 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Open reading frames ; Database ; Genetic nomenclature ; Codon bias ; Duplicated genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The amount of nucleotide sequence data is increasing exponentially. We therefore continued our effort to make a comprehensive database for the yeast Saccharomyces cerevisiae. In this database (ListA2) we have compiled 1001 protein coding sequences from this organism. Each sequence has been attributed a single genetic name and in the case of allelic duplicated sequences, synonyms are given, if necessary. For the nomenclature we have introduced a standard principle for naming gene sequences based on priority rules. We have also applied a simple method to distinguish duplicated sequences of one and the same gene from non-allelic sequences of duplicated genes. By using these principles we have sorted out a lot of confusion in the literature and databanks. Along with the genetic name, the mnemonic from the EMBL databank, the codon bias, reference of the publication of the sequence and the EMBL accession numbers are included for each entry. The database is available on request.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00336752
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  • 3
    Electronic Resource
    Electronic Resource
    Mitochondrial and nuclear mitoribosomal suppressors that enable misreading of ochre codons in yeast mitochondria (1984)
    Springer
    Current genetics 9 (1984), S. 11-19 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondrial ochre mutations ; Specificity of suppressors ; Mitoribosomal misreading ; Intron-encoded maturases
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We describe studies on the action spectra of the mitochondrial suppressor mim3-1 and the three alleles of nuclear suppressor nam3. Their specificity of action was tested on 516 mit − mutations located in different mitochondrial genes. The degree of suppression was quantified by the extent of cytochrome oxidase and cytochrome b synthesis. We show that the four suppressors are allele-specific gene-nonspecific informational suppressors. They would act by changing the structure of the small mitoribosomal subunit which would decrease fidelity of translation enabling misreading of some but not all ochre codons. The implications of the results on the role of intron encoded maturases are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00396199
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  • 4
    Electronic Resource
    Electronic Resource
    Mitochondrial and nuclear mitoribosomal suppressors that enable misreading of ochre codons in yeast mitochondria (1984)
    Springer
    Current genetics 9 (1984), S. 1-10 
    Details
    ISSN: 1432-0983
    Keywords: Yeast ; Mitochondrial ochre mutations ; Nuclear informational suppressors ; Mitochondrial informational suppressors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A systematic search for suppressors of mutations which cause a deficiency in the splicing of mitochondrial RNA has been undertaken. These splicing mutations were localized in the mRNA-maturase coding sequence of the second intron of the cob-boxgene, i.e. in the box3locus. A total of 953 revertants (mostly spontaneous in origin) were isolated and their genetic nature (nuclear vs. mitochondrial) and phenotype characterized. Most revertants were mitochondrially determined and displayed a wild-type phenotype. A mitochondrial suppressor unlinked with the box3 −target mutation was uncovered among the revertants displaying a pseudo-wild phenotype: out of 26 revertants analyzed, derived from 7 different box3− mutants only one such suppressor mutation mim3-1 was found. It was localized by rho− deletion mapping in the region between the oxi2 and oxi3 gene, within (or in the vicinity) the gene specifying the 15S ribosomal RNA. Nuclear suppressors were isolated from seven different box3 −mutants. All were recessive and had a pseudo-wild phenotype. Three such suppressors nam3-1, nam3-2 and nam3-3 were investigated more extensively. Tetrad analysis has shown that they are alleles of the same nuclear locus NAM3 and mitotic analysis has shown that they do not segregate mitotically.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00396198
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  • 5
    Electronic Resource
    Electronic Resource
    The CBP2 gene from Saccharomyces douglasii is a functional homologue of the Saccharomyces cerevisiae gene and is essential for respiratory growth in the presence of a wild-type (intron-containing) mitochondrial genome (1996)
    Springer
    Molecular genetics and genomics 250 (1996), S. 316-322 
    Details
    ISSN: 1617-4623
    Keywords: Key words Saccharomyces douglasii ; Saccharomyces cerevisiae ; CBP2 ; Mitochondria ; Pre-mRNA processing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  In Saccharomyces cerevisiae the only known role of the CBP2 gene is the excision of the fifth intron of the mitochondrial cyt b gene (bI5). We have cloned the CBP2 gene from Saccharomyces douglasii (a close relative of S. cerevisiae). A comparison of the S. douglasii and S. cerevisiae sequences shows that there are 14% nucleotide substitutions in the coding region, with transitions being three times more frequent than transversions. At the protein level sequence identity is 87%. We have demonstrated that the S. douglasii CBP2 gene is essential for respiratory growth in the presence of a wild-type S. douglasii mitochondrial genome, but not in the presence of an intronless S. cerevisiae mitochondrial genome. Also the S. douglasii and S. cerevisiae CBP2 genes are completely interchangeable, even though the intron bI5 is absent from the S. douglasii mitochondrial genome.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02174389
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  • 6
    Electronic Resource
    Electronic Resource
    Novel class of nuclear genes involved in both mRNA splicing and protein synthesis in Saccharomyces cerevisiae mitochondria (1989)
    Springer
    Molecular genetics and genomics 215 (1989), S. 517-528 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; Nuclear genes ; Mitochondrial translation ; Mitochondrial splicing ; Suppression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have cloned three distinct nuclear genes, NAM1, NAM7, and NAM8, which alleviate mitochondrial intron mutations of the cytochrome b and COXI (subunit I of cytochrome oxidase) genes when present on multicopy plasmids. These nuclear genes show no sequence homology to each other and are localized on different chromosomes: NAM1 on chromosome IV, NAM7 on chromosome XIII and NAM8 on chromosome VIII. Sequence analysis of the NAM1 gene shows that it encodes a protein of 440 amino acids with a typical presequence that would target the protein to the mitochondrial matrix. Inactivation of the NAM1 gene by gene transplacement leads to a dramatic reduction of the overall synthesis of mitochondrial protein, and a complete absence of the COXI protein which is the result of a specific block in COXI pre-mRNA splicing. The possible mechanisms by which the NAM1 gene product may function are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00427051
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  • 7
    Electronic Resource
    Electronic Resource
    Divergence of the mitochondrial leucyl tRNA synthetase genes in two closely related yeasts Saccharomyces cerevisiae and Saccharomyces douglasii: A paradigm of incipient evolution (1988)
    Springer
    Molecular genetics and genomics 213 (1988), S. 297-309 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; Mitochondria ; pre-mRNA splicing ; tRNA synthetase gene ; Incipient evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We studied the NAM2 genes of Saccharomyces douglasii and Saccharomyces cerevisiae, and showed that they are interchangeable for all the known functions of these genes, both mitochondrial protein synthesis and mitochondrial mRNA splicing. This confirms the prediction that the S. douglasii NAM2D gene encodes the mitochondrial leucyl tRNA synthetase (EC 6.1.1.4). The observation that these enzymes are interchangeable for their mRNA splicing functions, even though there are significant differences in the intron/exon structure of their mitochondrial genome, suggests that they may have a general role in yeast mitochondrial RNA splicing. A short open reading frame (ORF) precedes the synthetase-encoding ORF, and we showed that at least in S. cerevisiae this is not essential for the expression of the gene; however, it may be involved in a more subtle type of regulation. Sequence comparisons of S. douglasii and S. cerevisiae revealed a particularly interesting situation from the evolutionary point of view. It appears that the two yeasts have diverged relatively recently: there is remarkable nucleotide sequence conservation, with no deletions or insertions, but numerous (albeit non-saturating) silent substitutions resulting from transitions. This applies not only to the NAM2 coding regions, but also to two other ORFs flanking the NAM2 ORF. The regions between the ORFs (believed to be intergenic regions) are much less conserved, with several deletions and insertions. Thus S. douglasii and S. cerevisiae provide an ideal system for the study of molecular evolution, being two yeasts “caught in the act” of speciation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00339595
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  • 8
    Electronic Resource
    Electronic Resource
    Mitochondrial dysfunction in yeast expressing the cytoplasmic male sterility T-urf13 gene from maize: analysis at the population and individual cell level (1993)
    Springer
    Molecular genetics and genomics 236 (1993), S. 299-308 
    Details
    ISSN: 1617-4623
    Keywords: Texas maize cytoplasmic male sterility ; Saccharomyces cerevisiae ; Mitochondria ; Image and flow cytometry ; 3,3′-Dihexyloxacarbocyanine iodide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The urf13TW gene, which is derived from the mitochondrial T-urf13 gene responsible for Texas cytoplasmic male sterility in maize, was expressed in Saccharomyces cerevisiae by targeting its translation product into mitochondria. Analysis by oxygraphy at the population level revealed that in the presence of methomyl the oxygen uptake of intact yeast cells carrying the targeted protein is strongly stimulated only with ethanol as respiratory substrate and not with glycerol, lactate, pyruvate, or acetate. When malate is the substrate oxidized by isolated mitochondria, interaction between the targeted protein and methomyl results in significant inhibition of oxygen uptake. This inhibition is eliminated and oxygen uptake is stimulated by subsequent addition of NAD+. Using 3,3′-dihexyloxacarbocyanine iodide [DiOC6(3)] as probe, interactive laser scanning and flow cytometry, which permit analysis at the individual cell level, demonstrated that specific staining of the mitochondrial compartment is obtained and that DiOC6(3) fluorescence serves as a measure of the membrane potential. Finally, it was shown that, as in T cytoplasm maize mitochondria, HmT toxin and methomyl dissipate the membrane potential of yeast mitochondria that carry the foreign protein. Furthermore, the results suggest that the HmT toxin and methomyl response is related to the plasmid copy number per cell and that the deleterious effect induced by HmT toxin is stronger than that of methomyl.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00277126
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  • 9
    Electronic Resource
    Electronic Resource
    The NAM1/MTF2 nuclear gene product is selectively required for the stability and/or processing of mitochondrial transcripts of the atp6 and of the mosaic, cox1 and cytb genes in Saccharomyces cerevisiae (1993)
    Springer
    Molecular genetics and genomics 240 (1993), S. 419-427 
    Details
    ISSN: 1617-4623
    Keywords: Yeast ; Nucleo-mitochondrial intraction ; RNA processing ; RNA stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The NAM1/MTF2 gene was firstly isolated as a multicopy suppressor of mitochondrial splicing deficiencies and independently as a gene of which a thermosensitive allele affects mitochondrial transcription in organello. To determine which step in mitochondrial RNA metabolism is controlled in vivo by the NAM1 gene, mitochondrial transcripts of seven transcription units from strains carrying an inactive nam1::URA3 gene disruption in various mitochondrial genetic backgrounds were analysed by Northern blot hybridisations. In a strain carrying an intron-containing mitochondrial genome, the inactivation of the NAM1 gene led to a strong decrease in (or total absence of) the mosaic cytb and cox1 mRNAs and in transcripts of the atp6-rf3/ens2 genes, which are co-transcribed with cox1. Neither the accumulation of unspliced cytb or cox1 pre-mRNAs, nor that of excised circular intron molecules of ai1 or ai2 were observed, but the abundance of the bi1 and ai7 lariats was comparable to that observed in the wild-type strain, thus demonstrating that transcription of the cytb and cox1 genes does occur. In strains carrying the intron-less mitochondrial genome with or without the rf3/ens2 sequence, wild-type amounts of cytb and cox1 mRNAs were detected while the amount of the atp6 mRNA was always strongly decreased. The abundance of transcripts from five other genes was either slightly (21S rRNA) or not at all (cox2, cox3, atp9 and 15S rRNA) affected by the nam1 inactivation. This analysis leads to the conclusion that the NAM1 protein is not a general mitochondrial transcription factor, but rather is predominantly and selectively required for the processing and/or for the stability of cytb and cox1 intron-containing pre-mRNAs and of the atp6 transcripts. Since the original intronic mutations suppressed by the amplification of the NAM1 gene are situated in stem-loop rich structures, we propose that the NAM1 protein is a stem-loop RNA-binding protein that plays a role in determining RNA stability.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00280396
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  • 10
    Electronic Resource
    Electronic Resource
    III. Yeast sequencing reports. The complete sequence of the unit YCR59, situated between CRY1 and MAT, reveals two long open reading frames, which cover 91% of the 10·1 kb segment (1991)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 7 (1991), S. 413-424 
    Details
    ISSN: 0749-503X
    Keywords: Yeast ; Chromosome III ; sequence ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have entirely sequenced YCR59, which is a 10·1 kb segment of the right arm of chromosome III, and is a part of the clone E5F from the Newlon collection. The segment contains two long open reading frames (ORFs): YCR591 which starts in the adjacent fragment H9G (situated towards CRY1 and the centromere), and continues with 1833 codons in YCR59. The second ORF YCR592 is 1226 codons long and encoded entirely within YCR59. The two ORFs represent 91% of the total length of the segment. Excellent agreement in both location and length is found between the ORFs YCR591 and YCR592 and the transcripts 86 and 87 respectively in the Yoshikawa and Isono (1990) map of chromosome III. The two ORFs correspond to new genes and show no significant similarity with any known genes.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320070411
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