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In vitro and in vivo studies on the mitochondrial import of CBS1, a translational activator of cytochrome b in yeast

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Summary

Translation of mitochondrial cytochrome b mRNA in yeast is activated by the product of the nuclear gene CBS1. CBS1 encodes a 27 kDa precursor protein, which is cleaved to a 24 kDa mature protein during the import into isolated mitochondria. The sequences required for mitochondrial import reside in the amino-terminal end of the CBS1 precursor. Deletion of the 76 amino-terminal amino acids renders the protein incompetent for mitochondrial import in vitro and non-functional in vivo. When present on a high copy number plasmid and under the control of a strong yeast promoter, biological function can be restored by this truncated derivative. This observation indicates that the CBS1 protein devoid of mitochondrial targeting sequences can enter mitochondria in vivo, possibly due to a bypass of the mitochondrial import system.

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References

  • Ammerer G (1983) Expression of genes in yeast using the ADC1 promoter. Methods Enzymol 101:192–201

    Google Scholar 

  • Banroques J, Perea J, Jacq C (1987) Efficient splicing of two yeast mitochondrial introns controlled by a nuclear-encoded maturase. EMBO J 6:1085–1091

    Google Scholar 

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

    Google Scholar 

  • Costanzo MC, Fox TD (1988) Specific translational activation by nuclear gene products occurs in the untranslated leader of a yeast mitochondrial mRNA. Proc Natl Acad Sci USA 85:2677–2681

    Google Scholar 

  • 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 5:3637–3641

    Google Scholar 

  • Daum G, Böhni PC, Schatz G (1982) Import of proteins into mitochondria: cytochrome b 2and cytochrome c peroxidase are located in the intermembrane space of yeast mitochondria. J Biol Chem 257:13028–13033

    Google Scholar 

  • 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 260:1513–1520

    Google Scholar 

  • Douglas MG, McCammon MT, Vassarotti A (1986) Targeting proteins into mitochondria. Microbiol Rev 50:166–178

    Google Scholar 

  • Forsbach V, Rödel G (1987) Primary structure of the yeast CBS1 gene product, necessary to activate translation of mitochondrial COB mRNA. In: Papa S, Chance B, Ernster L (eds) Cytochrome systems. Plenum Press, New York, London, pp 169–176

    Google Scholar 

  • Fox TD (1986) Nuclear gene products required for translation of specific mitochondrially coded mRNAs in yeast. Trends Genet 2:97–100

    Google Scholar 

  • Fujiki Y, Hubbard AL, Fowlern S, Lazarow PB (1982) Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum. J Cell Biol 93:97–102

    Google Scholar 

  • Hartl F-U, Schmidt B, Wachter E, Weiss H, Neupert W (1986) Transport into mitochondria and intramitochondrial sorting of the Fe/S protein of ubiquinol-cytochrome c reductase. Cell 47:939–951

    Google Scholar 

  • 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 11:185–191

    Google Scholar 

  • Kuo C, Campbell JL (1983) Cloning of Saccharomyces cerevisiae DNA replication genes: Isolation of the CDC8 gene and two genes that compensate for the cdc8-1 mutation. Mol Cell Biol 3:1730–1737

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Lütcke HA, Chow KC, Mickel FS, Moss KA, Kern HF, Scheele GA (1987) Selection of AUG initiation codons differs in plants and animals. EMBO J 6:43–48

    Google Scholar 

  • 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 258:9459–9468

    Google Scholar 

  • Myers AM, Tzagoloff A (1986) Genetics of mitochondrial biogenesis. Annu Rev Biochem 55:249–285

    Google Scholar 

  • Pfanner N, Hoeben P, Tropschug M, Neupert W (1987) The carboxyl-terminal two-thirds of the ADP/ATP carrier polypeptide contains sufficient information to direct translocation into mitochondria. J Biol Chem 262:14851–14854

    Google Scholar 

  • Pfanner N, Hartl F-U, Neupert W (1988a) Import of proteins into mitochondria: a multi-step process. Eur J Biochem 175:205–212

    Google Scholar 

  • Pfanner N, Pfaller R, Neupert W (1988b) How finicky is mitochondrial protein import? Trends Biochem Sci 13:165–167

    Google Scholar 

  • Poutre CG, Fox TD (1987) PET111, a Saccharomyces cerevisiae nuclear gene required for translation of the mitochondrial mRNA encoding cytochrome oxidase subunit II. Genetics 115:637–647

    Google Scholar 

  • Rao JKM, Argos P (1986) A conformational preference parameter to predict helices in integral membrane proteins. Biochim Biophys Acta 869:197–214

    Google Scholar 

  • 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 11:41–45

    Google Scholar 

  • 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 206:45–50

    Google Scholar 

  • Rödel G, Michaelis U, Forsbach V, Kreike J, Kaudewitz F (1986) Molecular cloning of the yeast nuclear genes CBS1 and CBS2. Curr Genet 11:47–53

    Google Scholar 

  • 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 9:641–648

    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 

  • Schatz G (1987) Signals guiding proteins to their correct locations in mitochondria. Eur J Biochem 165:1–6

    Google Scholar 

  • Schulze M, Rödel G (1988) SCO1, a yeast nuclear gene essential for accumulation of mitochondrial cytochrome c oxidase subunit II. Mol Gen Genet 211:492–498

    Google Scholar 

  • Schulze M, Rödel G (1989) Accumulation of the cytochrome c oxidase subunits I and II in yeast requires a mitochondrial membrane-associated protein, encoded by the nuclear SCO1 gene. Mol Gen Genet (in press)

  • Sherman F, Fink GR, Lawrence CW (1986) Methods in Yeast Genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Strick CA, Fox TD (1987) Saccharomyces cerevisiae positive regulatory gene PET111 encodes a mitochondrial protein that is translated from a mRNA with a long 5′-leader. Mol Cell Biol 7:2728–2734

    Google Scholar 

  • Stuart RA, Neupert W, Tropschug M (1987) Deficiency in mRNA splicing in a cytochrome c mutant of Neurospora crassa: importance of carboxy terminus for import of apocytochrome c into mitochondria. EMBO J 6:2131–2137

    Google Scholar 

  • Von Heijne G (1986) Mitochondrial targeting sequences may form amphiphilic helices. EMBO J 5:1335–1342

    Google Scholar 

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Communicated by D.M. Lonsdale

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Körte, A., Forsbach, V., Gottenöf, T. et al. In vitro and in vivo studies on the mitochondrial import of CBS1, a translational activator of cytochrome b in yeast. Mol Gen Genet 217, 162–167 (1989). https://doi.org/10.1007/BF00330956

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  • DOI: https://doi.org/10.1007/BF00330956

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