Advertisement

Current Genetics

, Volume 26, Issue 1, pp 15–20 | Cite as

ERV1 is involved in the cell-division cycle and the maintenance of mitochondrial genomes in Saccharomyces cerevisiae

  • Thomas Lisowsky
Original Articles

Abstract

In former studies it was found that the ERV1 gene is essential for cell viability and for the biogenesis of functional mitochondria. A temperature-sensitive nuclear mutant exhibits a severe reduction in all the mitochondrial transcripts. Elimination of the gene leads to growth arrest after a few cell divisions. The putative gene product bears the characteristics of a regulatory factor since it has low expression rate and a high content of charged amino acids. In this study it is further verified that the ERV1 gene alone is responsible for the observed cellular and mitochondrial defects. The 5′ region of the gene is analysed by DNA deletions and complementation studies. Expression of the gene under the control of the GAL1-10 promoter in a disruption strain of ERV1 allows a more detailed specification of its influence on mitochondrial and cellular functions. Immediate and complete loss of mitochondrial genomes is observed after the promoter has been shut off, whereas the yeast cells are still able to grow for a limited time under these conditions. Analysis of the cells by in-vivo DNA flurorescence demonstrates a specific arrest in the cell-division cycle as the terminal phenotype. To further characterize the temperature-sensitive allele of ERV1 the mutated gene has been isolated and sequenced. A single point mutation which leads to the exchange of a single amino acid is found in the reading frame.

Key words

Cell-division cycle Mitochondrial genome Nuclear mutation Saccharomyces cerevisiae 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Attardi G, Schatz G (1988) Biogenesis of mitochondria. Annu Rev Cell Biol 4:289–333Google Scholar
  2. Baker KP, Schatz G (1991) Mitochondrial proteins essential for viability mediate protein import into yeast mitochondria. Nature 349:205–208Google Scholar
  3. 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 80:3963–3965Google Scholar
  4. Birnboim HC, Doly JD (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523Google Scholar
  5. Broach JR, Pringle JR, Jones EW (eds) (1991) The molecular and cellular biology of the yeast Sacaromyces: I. Genome dynamics, protein synthesis and energetics. II. Gene expression. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  6. Campbell JL, Newlon CS (1991) Chromosomal DNA replication. In: Broach JR, Pringle JR, Jones EW (eds) The molecular and cellular biology of the yeast Saccharomyces: Genome dynamics, protein synthesis and energetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 109–146Google Scholar
  7. Chen X-J, Guan M-X, Clark-Walker GD (1993) MGM1, a nuclear gene involved in maintenance of the mitochondrial genome in Saccharomyces cerevisiae. Nucleic Acids Res 21:3473–3477Google Scholar
  8. Diffley JFX, Stillman B (1991) A close relative of the nuclear, chromosomal high-mobility-group protein HMG1 in yeast mitochondria. Proc Natl Acad Sci USA 88:7864–7868Google Scholar
  9. Elledge SJ, Davis RW (1987) Identification and isolation of the gene encoding the small subunit of ribonucleotide reductase from Saccharomyces cerevisiae: DNA-damage-inducible gene required for mitotic viability. Mol Cell Biol 7:2783–2793Google Scholar
  10. Grivell LA (1989) Nucleo-mitochondrial interactions in yeast mitochondrial biogenesis. Eur J Biochem 182:477–493Google Scholar
  11. Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids J Mol Biol 166:557–580Google Scholar
  12. Hartwell LH, Culotti J, Reid B (1970) Genetic control of the celldivision cycle in yeast. I. detection of mutants. Proc Natl Acad Sci USA 66:352–359Google Scholar
  13. Hill JE, Myers AM, Koerner TJ, Tzagoloff A (1986) Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast 2:163–167Google Scholar
  14. Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells with alkali cations. J Bacteriol 153:163–168Google Scholar
  15. Jang SH, Jaehning JA (1991) The yeast mitochondrial RNA polymerase specificity factor, MTF1, is similar to bacterial σ factors. J Biol Chem 266:22671–22677Google Scholar
  16. Janitor M, Subik J (1993) Molecular cloning of the PEL1 gene of Saccharomyces cerevisiae that is essential for viability of petite mutants. Curr Genet 24:307–312Google Scholar
  17. Johnston M, Davis RW (1984) Sequences that regulate the divergent GAL1-10 promoter in Saccharomyces cerevisiae. Mol Cell Biol 4:1440–1448Google Scholar
  18. Lisowsky T (1992) Dual function of a new nuclear gene for oxidative phosphorylation and vegetative growth in yeast. Mol Gen Genet 232:58–64Google Scholar
  19. Lisowsky T (1993) A high copy number of yeast γ-glutamylcysteine synthetase suppresses a nuclear mutation affecting mitochondrial translation. Curr Genet 23:408–413Google Scholar
  20. Lisowsky T, Michaelis G (1988) A nuclear gene essential for mitochondrial replication suppresses a defect of mitochondrial transcription in Saccharomyces cerevisiae. Mol Gen Genet 214:218–223Google Scholar
  21. 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 219:125–128Google Scholar
  22. Masters BS, Stohl LL, Clayton DA (1987) Yeast mitochondrial RNA polymerase is homologous to those encoded by bacteriophage T3 and T7. Cell 51:89–99Google Scholar
  23. Merill BM, William KR, Chase JW, Konigsberg WH (1984) Photochemical crosslink of the Escherichia coli single-stranded DNA-binding protein to oligodeoxynucleotides: identification of phenylalanine 60 as the site of crosslinking. J Biol Chem 259:10850–10856Google Scholar
  24. Michaelis C, Mannhaupt G, Pratje E, Fisher 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, Cold Spring Harbor, New York, pp 311–321Google Scholar
  25. Nakai M, Endo T, Hase T, Matsubara H (1993) Intramitochondrial protein sorting-isolation and characterization of the yeast MSP1 gene which belongs to a novel family of putative ATPases. J Biol Chem 268:24262–24269Google Scholar
  26. Newlon CS (1988) Yeast chromosome replication and segregation. Microbiol Rev 52:568–601Google Scholar
  27. Newlon CS, Ludescher RD, Walter SK (1979) Production of petites by cell cycle mutants of Saccharomyces cerevisiae defective in DNA synthesis. Mol Gen Genet 169:189–194Google Scholar
  28. Parisi MA, Xu B, Clayton DA (1993) A human mitochondrial transcriptional activator can functionally replace a yeast mitochondrial HMG-box protein both in vivo and in vitro. Mol Cell Biol 13:1951–1961Google Scholar
  29. Pringle JR, Hartwell LH (1981) Saccharomyces cerevisiae cell cycle. In: Strathern JN, Jones EW, Broach JR (eds) The molecular biology of the yeast Saccharomyces: Life cycle and inheritance. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 97–142Google Scholar
  30. Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101:202–211Google Scholar
  31. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  32. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467Google Scholar
  33. Tzagoloff A (1982) Mitochondria. In: Siekevitz P (ed) Cellular organelles. Plenum, New York, pp 235–322Google Scholar
  34. Tzagoloff A, Dieckmann CL (1990) PET genes of Saccharomyces cerevisiae. Microbiol Rev 54:211–225Google Scholar
  35. Williamson DH, Fennell DJ (1975) The use of fluorescent DNA-binding agents for detecting and separating yeast mitochondrial DNA. Methods Cell Biol 17:335–351Google Scholar
  36. 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 33:103–119Google Scholar
  37. Ziaja K, Michaelis G, Lisowsky T (1993) Nuclear control of messenger RNA expression for mitochondrial ATPase subunit 9 in a new yeast mutant. J Mol Biol 229:909–916Google Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Thomas Lisowsky
    • 1
  1. 1.Botanisches InstitutHeinrich-Heine Universität DüsseldorfDüsseldorfGermany

Personalised recommendations