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.
Similar content being viewed by others
References
Attardi G, Schatz G (1988) Biogenesis of mitochondria. Annu Rev Cell Biol 4:289–333
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
Baker KP, Schatz G (1991) Mitochondrial proteins essential for viability mediate protein import into yeast mitochondria. Nature 349:205–208
Bennetzen JL, Hall BD (1982) Codon selection in yeast. J Biol Chem 257:3026–3031
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
Birnboim HC, Doly JD (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
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
Clayton DA (1984) Transcription of the mammalian mitochondrial genome. Annu Rev Biochem 53:573–594
Drlica K, Rouviere-Yanic J (1987) Histone-like proteins of bacteria. Microbiol Rev 51:301–319
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
Fisher RP, Lisowsky T, Breen GAM, Clayton DA (1991) A rapid, efficient method for purifying DNA-binding proteins. J Biol Chem 266:9153–9160
Grivell LA (1989) Nucleo-mitochondrial interactions in yeast mitochondrial biogenesis. Eur J Biochem 182:477–493
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
Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580
Hartl F-U, Pfanner N, Donald WN, Neupert W (1989) Mitochondrial protein import. Biochim Biophys Acta 988:1–45
Hill JE, Myers AM, Koerner TJ, Tzagoloff A (1986) Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast 2:163–167
Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells with alkali cations. J Bacteriol 153:163–168
Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132
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
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
Lisowsky T, Schweizer E, Michaelis G (1987) A nuclear mutation affecting mitochondrial transcription in Saccharomyces cerevisiae. Eur J Biochem 164:559–563
Mayes ELV (1982) High mobility group proteins. In: John EW (ed) The HMG chromosomal proteins. Academic Press, London, pp 9–40
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
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
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
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
Parisi MA, Clayton DA (1991) Similarity of human mitochondrial transcription factor 1 to high mobility group proteins. Science 252:965–969
Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
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
Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101:202–211
Tzagoloff A (1982) Mitochondria. In: Siekevitz P (ed) Cellular organelles. Plenum, New York, pp 41–59
Tzagoloff A, Dieckmann CL (1990) PET genes of Saccharomyces cerevisiae. Microbiol Rev 54:211–225
Vener K, Schatz G (1988) Protein translocation across membranes. Science 241:1307–1313
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
Author information
Authors and Affiliations
Additional information
Communicated by W. Gajewski
Rights and permissions
About this article
Cite this article
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
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00299137