Molecular and General Genetics MGG

, Volume 230, Issue 1–2, pp 177–185

Association of cytochrome b translational activator proteins with the mitochondrial membrane: implications for cytochrome b expression in yeast

  • Uwe Michaelis
  • Andreas Körte
  • Gerhard Rödel
Article

Summary

The products of the nuclear genes CBS1 and CBS2 are both required for translational activation of mitochondrial apocytochrome b in yeast. We report the intramitochondrial localization of both proteins by use of specific antisera. Based on its solubilization properties the CBS1 protein is presumed to be a component of the mitochondrial membrane; the detergent concentrations needed to release CBS1 from mitochondria are almost the same as for cytochrome c1. In contrast, CBS2 behaves like a soluble protein, with some characteristics of a membrane-associated protein. A model is presented for translational activation of cytochrome b, which might also be applicable to translational regulation of other mitochondrial genes.

Key words

Translational activation Cytochrome b Mitochondria Yeast CRS1 CBS2 

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References

  1. Ackerman SH, Gatti DL, Gellefors P, Douglas MG, Tzagoloff A (1991) ATP13, a nuclear gene of Saccharomyces cerevisiae essential for the expression of subunit 9 of the mitochondrial ATPase. FEBS Lett 278:234–238Google Scholar
  2. Attardi G, Schatz G (1988) Biogenesis of mitochondria. Annu Rev Cell Biol 4:289–333Google Scholar
  3. Costanzo MC, Fox TD (1986) Product of Saccharomyces cerevisiae nuclear gene PET494 activates translation of a specific mitochondrial mRNA. Mol Cell Biol 6:3694–3703Google Scholar
  4. Costanzo MC, Fox TD (1988) Specific translational activation by nuclear gene products occurs in the 5′ untranslated leader of a yeast mitochondrial mRNA. Proc Natl Acad Sci USA 85:2677–2681Google Scholar
  5. Costanzo MC, Fox TD (1990) Control of mitochondrial gene expression in Saccharomyces cerevisiae. Annu Rev Genet 24:91–113Google Scholar
  6. Costanzo MC, Seaver EC, Fox TD (1989) The PET54 gene of Saccharomyces cerevisiae: Characterization of a nuclear gene encoding a mitochondrial translational activator and subcellular localization of its product. Genetics 122:297–305Google Scholar
  7. Dake E, Hofmann TJ, McIntire S, Hudson A, Zassenhaus HP (1988) Purification and properties of the major nuclease from mitochondria of Saccharomyces cerevisiae. J Biol Chem 263:7691–7702Google Scholar
  8. Decoster E, Simon M, Hatat D, Faye G (1990) The MSS51 gene product is required for the translation of the COX1 mRNA in yeast mitochondria. Mol Gen Genet 224:111–118Google Scholar
  9. De La Salle H, Jacq C, Slonimski PP (1982) Critical sequences within mitochondrial introns: Pleiotropic mRNA maturase and cis-dominant signals of the box intron controlling reductase and oxidase. Cell 28:721–732Google 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. Forsbach V, Pillar T, Gottenöf T, Rödel G (1989) Chromosomal localization and expression of CBS1, a translational activator of cytochrome b in yeast. Mol Gen Genet 218:57–63Google Scholar
  12. Forsburg SL, Guarente L (1989) Communication between mitochondria and the nucleus in regulation of cytochrome genes in the yeast Saccharomyces cerevisiae. Annu Rev Cell Biol 5:153–180Google Scholar
  13. Graak H, Grohmann L, Choli T (1988) Mitochondrial ribosomes of yeast: isolation of individual proteins and N-terminal sequencing. FEBS Lett 242:4–8Google Scholar
  14. Grivell LA (1989) Nucleo-mitochondrial interactions in yeast mitochondrial biogenesis. Eur J Biochem 182:477–493Google Scholar
  15. Haffter P, McMullin TW, Fox TD (1990) A genetic link between an mRNA-specific translational activator and the translation system in yeast mitochondria. Genetics 125:495–503Google Scholar
  16. Haffter P, McMullin TW, Fox TD (1991) Functional interactions among two yeast mitochondrial ribosomal proteins and an mRNA-specific translational activator. Genetics 127:319–326Google Scholar
  17. Hartl F-U, Pfanner N, Nicholson DW, Neupert W (1989) Mitochondrial protein import. Biochim Biophys Acta 988:1–45Google Scholar
  18. Hase T, Harabayashi M, Kawai K, Matsubara H (1987) A carboxy-terminal hydrophobic region of cytochrome c1 is necessary for functional assembly into complex III of the respiratory chain. J Biochem 102:411–419Google Scholar
  19. Körte A, Forsbach V, Gottenöf T, Rödel G (1989) In vitro and in vivo studies on the mitochondrial import of CBS1, a translational activator of cytochrome b in yeast. Mol Gen Genet 217:162–167Google Scholar
  20. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685Google Scholar
  21. Lamb MR, Anziano PQ, Glans KR, Hanson DK, Klapper HJ, Perlman PS, Mahler HR (1983) Functional domains in introns: RNA processing intermediates in cis- and trans-acting mutants in the penultimate intron of the mitochondrial gene for cytochrome b. J Biol Chem 258:1991–1999Google Scholar
  22. Marykwas DL, Fox TD (1989) Control of the Saccharomyces cerevisiae regulatory gene PET494: transcriptional repression by glucose and translational induction by oxygen. Mol Cell Biol 9:484–491Google Scholar
  23. McMullin TW, Haffter P, Fox TD (1990) A novel small-subunit ribosomal protein of yeast mitochondria that interacts functionally with an mRNA-specific translational activator. Mol Cell Biol 10:4590–4595Google Scholar
  24. Michaelis U, Rödel G (1990) Identification of CBS2 as a mitochondrial protein in Saccharomyces cerevisiae. Mol Gen Genet 223:394–400Google Scholar
  25. Muroff I, Tzagoloff A (1990) CBP7 codes for a co-factor required in conjunction with a mitochondrial maturase for splicing of its cognate intervening sequence. EMBO J 9:2765–2773Google Scholar
  26. Ohmen JD, Kloeckener-Gruissem B, McEwen JE (1988) Molecular cloning and nucleotide sequence of the PET122 gene required for expression of the mitochondrial COX3 gene in S. cerevisiae. Nucleic Acids Res 16:10783–10802Google Scholar
  27. Ooi BG, Lukins HB, Linnane AW, Nagley P (1987) Biogenesis of mitochondria: a mutation in the 5′-untranslated region of a yeast mitochondrial OLI1 mRNA leading to impairment in translation of subunit 9 of the mitochondrial ATPase complex. Nucleic Acids Res 15:1965–1977Google Scholar
  28. Papadopoulou B, Dekker P, Blom J, Grivell LA (1990) A 40 kd protein binds specifically to the 5′-untranslated regions of yeast mitochondrial mRNAs. EMBO J 9:4135–4143Google Scholar
  29. Penefsky HS, Tzagoloff A (1971) Extraction of water-soluble enzymes and proteins from membranes. Methods Enzymol 22:204–219Google Scholar
  30. Pillar T, Kreike J, Lang BF, Kaudewitz F (1983a) Nuclear mutants defective in processing of cob and oxi3 transcripts in Saccharomyces cerevisiae. In: Schweyen RJ, Wolf K, Kaudewitz F (eds) Mitochondria 1983. Walter de Gruyter, Berlin, pp 411–423Google Scholar
  31. Pillar T, Lang BF, Steinberger J, Vogt B, Kaudewitz F (1983b) Expression of the “split gene” cob in yeast mtDNA. J Biol Chem 258:7954–7959Google Scholar
  32. Poutre CG, Fox TD (1987) PET111, a Saccharomyces cerevisiae nuclear gene required for translation of the mitochondrial mRNA encoding cytochrome oxidase subunit II. Genetics 115:637–647Google Scholar
  33. Query CC, Bentley RC, Keene JD (1989) A common RNA recognition motif identified within a defined U1 RNA binding domain of the 70 kd U1 snRNP protein. Cell 57:89–101Google Scholar
  34. Rödel G (1986) Two yeast nuclear genes, CBS1 and CBS2, are required for translation of mitochondrial transcripts bearing the 5′-untranslated COB leader. Curr Genet 11:41–45Google Scholar
  35. 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
  36. 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
  37. 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
  38. Spithill TW, Trenbath MK, Lukins HB, Linnane AW (1978) Mutations of the mitochondrial DNA of Saccharomyces cerevisiae which affect the interaction between mitochondrial ribosomes and the inner mitochondrial membrane. Mol Gen Genet 164:155–162Google Scholar
  39. Weiss-Brummer B, Rödel G, Schweyen RJ, Kaudewitz F (1982) Expression of the split cob in yeast: evidence for a precursor of a “maturase” protein translated from intron 4 and preceding exons. Cell 29:527–5365Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • Uwe Michaelis
    • 1
  • Andreas Körte
    • 1
  • Gerhard Rödel
    • 2
  1. 1.Lehrstuhl GenetikUniversität MünchenMünchen 19Germany
  2. 2.Labor für Molekulare Biologic und Allgemeine PathologieInstitut für Pathologie der Universität UlmM üchen 45Germany

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