Molecular and General Genetics MGG

, Volume 211, Issue 3, pp 492–498

SCO1, a yeast nuclear gene essential for accumulation of mitochondrial cytochrome c oxidase subunit II

  • Marion Schulze
  • Gerhard Rödel


We have identified and isolated a novel yeast nuclear gene (SCO1) which is essential for accumulation of the mitochondrially synthesized subunit II of cytochrome c oxidase (CoxII). Analysis of the mitochondrial translation products in a sco1-1 mutant reveals a strong reduction in CoxII. Examination of mitochondrial transcripts by Northern blot hybridization shows that transcription and transcript maturation of OXI1, the gene coding for CoxII, is not affected. Therefore the SCO1 gene product must be involved in a post-transcriptional step in the synthesis of CoxII. We have isolated a 1.7 kb DNA fragment from a yeast gene bank which carries the functional SCO1 gene. Two RNA species of 0.9 kb and 1.2 kb, respectively, hybridize with this DNA fragment, which is localized on chromosome II. Cells whose chromosomal 1.7 kb fragment has been replaced by the yeast URA3 gene fail to accumulate CoxII and in addition subunit I of cytochrome c oxidase (CoxI). The possibility that the SCO1 gene product is bifunctional, i.e. required for both CoxI and CoxII accumulation, is discussed.

Key words

Cytochrome c oxidase Mitochondria PET genes Yeast 


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  1. Beggs JD (1978) Transformation of yeast by a replicating hybrid plasmid. Nature 275:104–108Google Scholar
  2. Birnboim H, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523Google Scholar
  3. Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S (1977) Construction and characterization of new cloning vehicles II. A multipurpose cloning system. Gene 2:95–113Google Scholar
  4. Carle GF, Olson MV (1985) An electrophoretic karyotype for yeast. Proc Natl Acad Sci USA 79:3756–3760Google Scholar
  5. Christianson T, Rabinowitz M (1983) Identification of multiple transcriptional initiation sites on the yeast mitochondrial genome by in vitro capping with guanylyltransferase. J Biol Chem 258:14025–14033Google Scholar
  6. Clewell DB (1972) Nature of Col1E 1 plasmid replication in E. coli in the presence of chloramphenicol. J Bacteriol 110:667–676Google Scholar
  7. Coruzzi G, Bonitz SG, Thalenfeld BE, Tzagoloff A (1981) Assembly of the mitochondrial membrane system. Analysis of the nucleotide sequence and transcripts in the oxil region of yeast mitochondrial DNA. J Biol Chem 256:12780–12787Google Scholar
  8. Costanzo MC, Fox TD (1986) Product of Saccharomyces cerevisiae nuclear gene PET494 activates translation of specific mitochondrial mRNA. Mol Cell Biol 6:3694–3703Google Scholar
  9. Costanzo MC, Seaver EC, Fox TD (1986) At least two nuclear gene products are specifically required for translation of a single yeast mitochondrial mRNA. EMBO J 5:3637–3641Google Scholar
  10. Dieckmann CL, Tzagoloff A (1985) Assembly of the mitochondrial membrane system: CBP6, a yeast nuclear gene necessary for synthesis of cytochrome b. J Biol Chem 260:1513–1520Google Scholar
  11. Dieckmann CL, Koerner TJ, Tzagoloff A (1984a) Assembly of the mitochondrial membrane system: CBP1, a yeast nuclear gene involved in 5′ end processing of cytochrome b pre mRNA. J Biol Chem 259:4722–4731Google Scholar
  12. Dieckmann CL, Homison G, Tzagoloff A (1984b) Assembly of the mitochondrial membrane system: Nucleotide sequence of a yeast nuclear gene (CBP1) involved in 5′ end processing of cytochrome b pre mRNA. J Biol Chem 259:4732–4738Google Scholar
  13. Douglas MG, Butow RA (1976) Variant forms of mitochondrial translation products in yeast: Evidence for location of determinants on mitochondrial DNA. Proc Natl Acad Sci USA 73:1083–1086Google Scholar
  14. Fasiolo F, Bonnet JH, Lacroute F (1981) Cloning of the yeast methionyl-tRNA synthetase gene. J Biol Chem 256:2324–2328Google Scholar
  15. Faye G, Simon M (1983) Analysis of a yeast nuclear gene involved in the maturation of mitochondrial pre-messenger RNA of the cytochrome oxidase subunit I. Cell 32:77–87Google Scholar
  16. Fox TD (1986) Nuclear gene products required for translation of specific mitochondrially coded mRNAs in yeast. Trends Genet 2:97–100Google Scholar
  17. Guarente L, Yocum RR, Gifford P (1982) A GAL10-CYC1 hybrid yeast promoter identifies the GALA regulatory region as an upstream site. Proc Natl Acad Sci USA 79:7410–7414Google Scholar
  18. Hill J, McGraw P, Tzagoloff A (1985) A mutation in yeast mitochondrial DNA results in a precise excision of the terminal intron of the cytochrome b gene. J Biol Chem 260:3235–3238Google Scholar
  19. Kloeckener-Gruissem B, McEwen JE, Poyton RO (1987) Nuclear functions required for cytochrome c oxidase biogenesis in Saccharomyces cerevisiae: Multiple trans-acting nuclear genes exert specific effects on expression of each of the cytochrome c oxidase subunits encoded on mitochondrial DNA. Curr Genet 12:311–322Google Scholar
  20. Koerner TJ, Hill J, Tzagoloff A (1985) Cloning and characterization of the yeast nuclear gene for subunit 5 of cytochrome oxidase. J Biol Chem 260:9513–9515Google Scholar
  21. Kreike J, Schulze M, Pillar T, Körte A, Rödel G (1986) Cloning of a nuclear gene MRS1 involved in the excision of a single group I intron (bI3) from the mitochondrial COB transcript in S. cerevisiae. Curr Genet 11:185–191Google Scholar
  22. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685Google Scholar
  23. Lang BF, Kaudewitz F (1982) Cytochrome c1-deficient mutants in Saccharomyces cerevisiae. Curr Genet 6:229–235Google Scholar
  24. Maarse AC, Van Loon APGM, Riezman H, Gregor I, Schatz G, Grivell LA (1984) Subunit IV of yeast cytochrome c oxidase: cloning and nucleotide sequencing of the gene and partial amino acid sequencing of the mature protein. EMBO J 3:2831–2837Google Scholar
  25. Mannhaupt G, Michaelis G, Pratje E, Schweizer E, Hawthorne DC (1983) A precursor to subunit II of cytochrome oxidase in Saccharomyces cerevisiae. In: Schweyen RJ, Wolf K, Kaudewitz F (eds) Mitochondrial 1983. W de Gruyter and Co, Berlin, New York, pp 449–454Google Scholar
  26. Mannhapt G, Beyreuther K, Michaelis G (1985) Cytochrome b, the var 1 protein, and subunits I and III of cytochrome c oxidase are synthesized without transient presequences in Saccharomyces cerevisiae. Eur J Biochem 150:435–439Google Scholar
  27. McEwen JE, Ko C, Kloeckner-Gruissem B, Poyton RO (1986) Nuclear functions required for cytochrome c oxidase biogenesis in Saccharomyces cerevisiae. J Biol Chem 261:11872–11879Google Scholar
  28. McGraw P, Tzagoloff A (1983) Assembly of the mitochondrial membrane system: Characterization of a yeast nuclear gene involved in the processing of the cytochrome b pre mRNA. J Biol Chem 258:9459–9468Google Scholar
  29. Michaelis G, Mannhaupt G, Pratje E, Fischer E, Naggert J, Schweizer E (1982) Mitochondrial translation products in nuclear respiration-deficient pet mutants of Saccharomyces cerevisiae. In: Attardi G, Borst P, Slonimski PP (eds) Mitochondrial genes. Cold Spring Harbor Laboratory. Cold Spring Harbor, New York, pp 311–321Google Scholar
  30. Nasmyth KA, Reed SI (1980) Isolation of genes by complementation in yeast: Molecular cloning of a cell-cycle gene. Proc Natl Acad Sci USA 77:2119–2123Google Scholar
  31. Needleman RB, Tzagoloff A (1975) Breakage of yeast: a method for processing multiple samples. Anal Biochem 64:545–549Google Scholar
  32. Patterson TE, Poyton RO (1986) COX8, the structural gene for yeast cytochrome c oxidase subunit VIII. J Biol Chem 261:17192–17197Google Scholar
  33. Pillar T, Lang BF, Steinberger J, Vogt B, Kaudewitz F (1983) Expression of the “split gene” cob in yeast mtDNA. J Biol Chem 258:7954–7959Google Scholar
  34. Poutre CG, Fox TD (1987) PET111, a Saccharomyces cerevisiae nuclear gene required for translation of the mitochondrial mRNA encoding oxidase subunit II. Genetics 115:637–647Google Scholar
  35. Pratje E, Guiard B (1986) One nuclear gene controls the removal of transient pre-sequences from the yeast proteins: one encoded by the nuclear the other by the mitochondrial genome. EMBO J 5:1313–1317Google Scholar
  36. Pratje E, Mannhaupt G, Michaelis G, Beyreuther K (1983) A nuclear mutation prevents processing of a mitochondrially encoded membrane protein in Saccharomyces cerevisiae. EMBO J 2:1049–1054Google Scholar
  37. Rigby PW, Dieckman M, Rhodes C, Berg P (1977) Labeling of deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251Google Scholar
  38. Roberts TM, Swanberg SL, Poteete A, Riedel G, Backman K (1980) A plamid cloning vehicle allowing a positive selection for inserted fragments. Gene 12:123–127Google Scholar
  39. Rödel G (1986) Two nuclear genes, CBS1 and CBS2, are required for translation of mitochondrial transcripts bearing the 5′-untranslated COB leader. Curr Genet 11:41–45Google Scholar
  40. Rödel G, Fox TD (1987) The yeast nuclear gene CBS1 is required for translation of mitochondrial mRNAs bearing the COB 5′ untranslated leader. Mol Gen Genet 206:45–50Google Scholar
  41. Rödel G, Körte A, Kaudewitz F (1985) Mitochondrial suppression of a yeast nuclear mutation which affects the translation of the mitochondrial apocytochrome b transcript. Curr Genet 9:641–648Google Scholar
  42. Rödel G, Michaelis U, Forsbach V, Kreike J, Kaudewitz F (1986) Molecular cloning of the yeast nuclear genes CBS1 and CBS2. Curr Genet 11:47–53Google Scholar
  43. Rose M, Grisafi P, Botstein D (1984) Structure and function of the yeast URA3 gene: expression in Escherichia coli. Gene 29:113–124Google Scholar
  44. Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101:202–211Google Scholar
  45. Seraphin B, Simon M, Faye G (1985) Primary structure of a gene for subunit V of the cytochrome c oxidase from Saccharomyces cerevisiae. Curr Genet 9:435–439Google Scholar
  46. Sherman F, Fink GR, Lawrence CW (1986) Methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  47. Simon M, Faye G (1984) Steps in processing of the mitochondrial cytochrome oxidase subunit I pre-mRNA affected by a nuclear mutation in yeast. Proc Natl Acad Sci USA 81:8–12Google Scholar
  48. Southern EM (1975) Detection of specific sequences among DNA fragments by gel electrophoresis. J Mol Biol 98:503–517Google Scholar
  49. St. John T, Davis R (1981) The organization and transcription of the galactose gene cluster of Saccharomyces. J Mol Biol 152:285–315Google Scholar
  50. Van Loon APGM, Van Eijk E, Grivell LA (1983) Biosynthesis of the ubiquinol-cytochrome c reductase complex in yeast. Discoordinate synthesis of the 11 kD subunit in response to increased gene copy number. EMBO J 2:1765–1770Google Scholar
  51. Wright RM, Ko C, Cumsky MG, Poyton RO (1984) Isolation and sequence of the structural gene for cytochrome c oxidase subunit VI from Saccharomyces cerevisiae. J Biol Chem 259:15401–15407Google Scholar
  52. Wright RM, Dircks LK, Poyton RO (1986) Characterization of COX9, the nuclear gene encoding the yeast mitochondrial protein cytochrome c oxidase subunit VIIa. J Biol Chem 261:17183–17191Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Marion Schulze
    • 1
  • Gerhard Rödel
    • 1
  1. 1.Institut für Genetik und Mikrobiologie der Universität MünchenMünchen 19Germany

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