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Mitochondrial protein import: isolation and characterization of the Saccharomyces cerevisiae MFT1 gene

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Summary

Mitochondrial targeting of an Atp2-LacZ fusion protein confers a respiration-defective phenotype on yeast cells. This effect has been utilized to select strains that grow on nonfermentable carbon sources, some of which have decreased levels of hybrid protein localized to the organelle. Many of the mutants obtained were also temperature-sensitive for growth on all media. The recessive mft (mitochondrial fusion targeting) mutants have been assigned to three complementation groups. MFT1 was cloned and sequenced: it encodes a 255 amino acid protein that is highly basic and has no predicted membrane-spanning domains or organelle-targeting sequences. The MFT1 gene is 91% identical to an open reading frame 3′ of the SIR3 gene. Evidence is presented that these two closely related genes could represent a recent gene duplication.

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References

  • Bankaitis VA, Johnson LM, Emr SD (1986) Isolation of yeast mutants defective in protein targeting to the vacuole. Proc Natl Acad Sci USA 83:9075–9079

    Google Scholar 

  • Bedwell DM, Klionsky DJ, Emr SD (1989) The yeast F1-ATPase β subunit precursor contains functionally redundant mitochondrial protein import information. Mol Cell Biol 7:4038–4047

    Google Scholar 

  • Benson SA, Hall MN, Silhavy TJ (1985) Genetic analysis of protein export in Escherichia coli K12. Annu Rev Biochem 54:101–134

    Google Scholar 

  • Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Chirico WJ, Waters MG, Blobel G (1988) 70K heat shock related proteins stimulate protein translocation into microsomes. Nature 332:805–810

    Google Scholar 

  • Deshaies RJ, Koch BD, Werner Washburne M, Craig E, Schekman R (1988) A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides. Nature 332:804–805

    Google Scholar 

  • Domdey H, Apostol B, Lin R-J, Newman A, Brody E, Abelson J (1984) Lariat structures are in vivo intermediates in yeast pre-mRNA splicing. Cell 39:611–621

    Google Scholar 

  • Douglas M, Geller B, Emr S (1984) Intracellular targeting and import of an F1-ATPase β-subunit-β-galactosidase hybrid protein into yeast mitochondria. Proc Natl Acad Sci USA 81:3983–3987

    Google Scholar 

  • Emr SD, Schauer I, Hansen W, Esmon P, Schekman R (1984) Invertase β-galactosidase hybrid proteins fail to be transported from the endoplasmic reticulum in Saccharomyces cerevisiae. Mol Cell Biol 4:2347–2355

    Google Scholar 

  • Emr SD, Vassarotti A, Garrett J, Geller BL, Takeda M, Douglas MG (1986) The amino terminus of the yeast F1-ATPase-β-subunit precursor functions as a mitochondrial import signal. J Cell Biol 102:523–533

    Google Scholar 

  • Gavel Y, Nilsson L, von Heijne G (1988) Mitochondrial targeting sequences. Why ‘non-amphiphilic’ peptides may still be amphiphilic. FEBS Lett 235:173–177

    Google Scholar 

  • Hurt E, Pesold-Hurt B, Schatz G (1984) The amino terminal region of an imported mitochondrial precursor polypeptide can direct cytoplasmic dihydrofolate reductase into the mitochondrial matrix. EMBO J 3:3149–3156

    Google Scholar 

  • Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press, New York, pp 1–506

    Google Scholar 

  • Miller J (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, New York, pp 352–355

    Google Scholar 

  • Ostermann J, Horwich AL, Neupert W, Hard FU (1989) Protein folding in mitochondria requires complex formation with hsp60 and ATP hydrolysis. Nature 341:125–130

    Google Scholar 

  • Pelham HRB (1986) Speculations on the functions of the major heat shock and glucose-regulated proteins. Cell 46:959–961

    Google Scholar 

  • Reid GA (1985) Transport of proteins into mitochondria. Curr Top Membr Transp 24:295–337

    Google Scholar 

  • Riezman H, Hay R, Witts C, Nelson N, Schatz G (1983) Yeast mitochondrial outer membrane specifically binds cytoplasmically synthesized precursors of mitochondrial proteins EMBO J 2:1113–1118

    Google Scholar 

  • Robinson JS, Klionsky DJ, Banta LM, Emr SD (1988) Protein sorting in Saccharomyces cerevisiae: Isolation of mutants defective in the delivery and processing of multiple vacuolar proteases. Mol Cell Biol 8:4939–4948

    Google Scholar 

  • Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101:202–211

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    CAS  PubMed  Google Scholar 

  • Sherman F, Fink GR, Hicks JB (1986) Methods in yeast genetics: A laboratory manual. Revised edn Cold Spring Harbor Laboratory Press, New York, pp 1–196

    Google Scholar 

  • Silhavy TJ, Shuman HA, Beckwith J, Schwartz M (1977) Use of gene fusions to study outer membrane protein localization in Escherichia coli. Proc Natl Acad Sci USA 74:5411–5415

    Google Scholar 

  • Sollner T, Griffiths G, Pfaller R, Planner N, Neupert W (1989) Mom19, an import receptor for mitochondrial precursor proteins. Cell 59:1061–1070

    Google Scholar 

  • Vestweber D, Brunner J, Baker A, Schatz G (1989) A 42K outermembrane protein is a component of the yeast mitochondrial protein import site. Nature 341:205–209

    Google Scholar 

  • Witte C, Jensen RE, Yaffe MP, Schatz G (1988) MAS1, a gene essential for yeast mitochondrial assembly, encodes a subunit of the mitochondrial processing protease.

  • Yaffe MP, Schatz G (1984) Two nuclear mutations that block mitochondrial protein import in yeast. Proc Natl Acad Sci USA 81:4819–4823

    Google Scholar 

  • Zwizinski C, Schleyer M, Neupert W (1984) Proteinaceous receptors for the import of mitochondrial precursor proteins. J Biol Chem 7850–7856

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Communicated by D.Y. Thomas

The sequence reported here has been listed in the EMBL Data Library with Accession Number X55360.

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Garrett, J.M., Singh, K.K., Vonder Haar, R.A. et al. Mitochondrial protein import: isolation and characterization of the Saccharomyces cerevisiae MFT1 gene. Molec. Gen. Genet. 225, 483–491 (1991). https://doi.org/10.1007/BF00261691

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  • DOI: https://doi.org/10.1007/BF00261691

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