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Dual function of a new nuclear gene for oxidative phosphorylation and vegetative growth in yeast

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Summary

A new gene essential for cell viability and indispensable for the biogenesis of a functional respiratory chain in Saccharomyces cerevisiae was isolated by complementing a temperature-sensitive mutant. This conditional nuclear mutation selectively affects oxidative phosphorylation at restrictive temperatures. At the molecular level a severe and complex defect inside mitochondria is observed, with drastically reduced levels of mitochondrial transcripts. Surprisingly a null mutation in this nuclear gene in a haploid yeast strain leads to cell death. Spores containing a disrupted copy of the gene exhibit a severe growth defect and cell division stops irreversibly after 3 to 4 days. It is shown that the null and conditional mutants are indeed allelic. This finding demonstrates a dual function of the gene product in oxidative phosphorylation and vegetative growth. The putative protein product, as deduced from the sequence of the relevant reading frame is characterized by a low molecular weight of approximately 14 kDa, a high content of charged amino acids and a very low codon bias index. A transcript of low abundance and with a length of about 600 nucleotides can be assigned to this gene.

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References

  • Attardi G, Schatz G (1988) Biogenesis of mitochondria. Annu Rev Cell Biol 4:289–333

    Google Scholar 

  • Aviv H, Leder P (1972) Purification of biologically active globin messenger RNA by chromatography on oligothymidylic-acid cellulose. Proc Natl Acad Sci USA 69:1408–1412

    Google Scholar 

  • Baker KP, Schatz G (1991) Mitochondrial proteins essential for viability mediate protein import into yeast mitochondria. Nature 349:205–208

    Google Scholar 

  • Bennetzen JL, Hall BD (1982) Codon selection in yeast. J Biol Chem 257:3026–3031

    Google Scholar 

  • Biggin MD, Gilson TJ, Hong GF (1983) Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci USA 80:3963–3965

    Google Scholar 

  • Birnboim HC, Doly JD (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523

    Google Scholar 

  • Broach JR, Strathern JN, Hicks JB (1979) Transformation in yeast: Development of a hybrid cloning vector and isolation of the CAN1 gene. Gene 8:121–133

    Google Scholar 

  • Clayton DA (1984) Transcription of the mammalian mitochondrial genome. Annu Rev Biochem 53:573–594

    Google Scholar 

  • Drlica K, Rouviere-Yanic J (1987) Histone-like proteins of bacteria. Microbiol Rev 51:301–319

    Google Scholar 

  • Elder RT, Loh EY, Davis RW (1983) RNA from the yeast transposable element TY1 has both ends in the direct repeats, a structure similar to retrovirus RNA. Proc Natl Acad Sci USA 80:2432–2436

    Google Scholar 

  • Fisher RP, Lisowsky T, Breen GAM, Clayton DA (1991) A rapid, efficient method for purifying DNA-binding proteins. J Biol Chem 266:9153–9160

    Google Scholar 

  • Grivell LA (1989) Nucleo-mitochondrial interactions in yeast mitochondrial biogenesis. Eur J Biochem 182:477–493

    Google Scholar 

  • Hamilton R, Watanabe CK, de Boer HA (1987) Compilation and comparison of the sequence context around AUG start codons in Saccharomyces cerevisiae mRNAs. Nucleic Acids Res 15:3581–3593

    Google Scholar 

  • Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580

    Google Scholar 

  • Hartl F-U, Pfanner N, Donald WN, Neupert W (1989) Mitochondrial protein import. Biochim Biophys Acta 988:1–45

    Google Scholar 

  • Hill JE, Myers AM, Koerner TJ, Tzagoloff A (1986) Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast 2:163–167

    Google Scholar 

  • Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells with alkali cations. J Bacteriol 153:163–168

    Google Scholar 

  • Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132

    Google Scholar 

  • Langford CJ, Gallwitz D (1983) Evidence for an intron-contained sequence required for the splicing of yeast RNA polymerase II transcripts. Cell 33:519–527

    Google Scholar 

  • Lisowsky T, Michaelis G (1989) Mutations in the genes for mitochondrial RNA polymerase and a second mitochondrial transcription factor of Saccharomyces cerevisiae. Mol Gen Genet 219:125–128

    Google Scholar 

  • Lisowsky T, Schweizer E, Michaelis G (1987) A nuclear mutation affecting mitochondrial transcription in Saccharomyces cerevisiae. Eur J Biochem 164:559–563

    Google Scholar 

  • Mayes ELV (1982) High mobility group proteins. In: John EW (ed) The HMG chromosomal proteins. Academic Press, London, pp 9–40

    Google Scholar 

  • Michaelis G, Mannhaupt G, Pratje E, Fischer E, Naggert J, Schweizer E (1982) Mitochondrial translation products in nuclear respiratory-deficient pet mutants of Saccharomyces cerevisiae. In: Slonimski P, Borst P, Attardi G (eds) Mitochondrial genes. Cold Spring Harbor Laboratory Press, New York, pp 311–321

    Google Scholar 

  • Najarian D, Dihanich ME, Martin N, Hopper AK (1987) DNA sequence and transcript mapping of MOD5: Features of the 5′ region which suggests two translational starts. Mol Cell Biol 7:185–191

    Google Scholar 

  • Nasmyth KA, Reed SI (1980) Isolation of genes by complementation in yeast: Molecular cloning of a cell cycle gene. Proc Natl Acad Sci USA 77:2119–2123

    Google Scholar 

  • Natsoulis G, Hilger F, Fink GR (1986) The HTS1 gene encodes both the cytoplasmic and mitochondrial histidine tRNA synthetase of S. cerevisiae. Cell 46:235–243

    Google Scholar 

  • Parisi MA, Clayton DA (1991) Similarity of human mitochondrial transcription factor 1 to high mobility group proteins. Science 252:965–969

    Google Scholar 

  • Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Sor F, Fukuhara H (1982) Identification of two erythromycin resistance mutations in the gene coding for the large ribosomal RNA in yeast. Nucleic Acids Res 10:6571–6577

    Google Scholar 

  • Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101:202–211

    Google Scholar 

  • Tzagoloff A (1982) Mitochondria. In: Siekevitz P (ed) Cellular organelles. Plenum, New York, pp 41–59

    Google Scholar 

  • Tzagoloff A, Dieckmann CL (1990) PET genes of Saccharomyces cerevisiae. Microbiol Rev 54:211–225

    Google Scholar 

  • Vener K, Schatz G (1988) Protein translocation across membranes. Science 241:1307–1313

    Google Scholar 

  • Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC vectors. Gene 33:103–119

    Google Scholar 

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Communicated by W. Gajewski

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Lisowsky, T. Dual function of a new nuclear gene for oxidative phosphorylation and vegetative growth in yeast. Molec. Gen. Genet. 232, 58–64 (1992). https://doi.org/10.1007/BF00299137

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