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Transcription of TIM9, a new factor required for the petite-positive phenotype of Saccharomyces cerevisiae, is defective in spt7 mutants

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Abstract.

TIM9 has been identified as an additional novel gene required for the petite-positive phenotype in Saccharomyces cerevisiae. tim9-1 was obtained through a screen for respiratory-deficient strains that are unable to survive in the absence of mitochondrial DNA. A point mutation found in the tim9-1 coding region converts codon 71 from Gly to Arg. Examination of genes encoding other Tim components indicated that the temperature-conditional alleles of essential genes for the viability of S. cerevisiae, TIM9, TIM10 and TIM12, are required for petite survival, while deletion of TIM8 and TIM13 has no notable effect on petite cell viability. Northern hybridization results suggested that the Spt7 transcription factor is strictly involved in transcription of TIM9 and that the synergistic lethality of tim9-1/spt7Δ dual mutations is due to the deficiency of TIM9 transcription together with defective function of the tim9-1 protein.

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References

  • Ackerman SH, Tzagoloff A (1990) Identification of two nuclear genes (ATP11, ATP12) required for assembly of the yeast F1-ATPase. Proc Natl Acad Sci USA 87:4986–4990

    Google Scholar 

  • Adam A, Endres M, Sirrenberg C, Lottspeich F, Neupert W, Brunner M (1999) Tim9, a new component of the TIM22.54 translocase in mitochondria. EMBO J 18:313–319

    Google Scholar 

  • Burke D, Dawson D, Stearns T (2000) Methods in yeast genetics: a cold spring harbor laboratory course manual 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.

    Google Scholar 

  • Chang SC, Heacock PN, Clancey CJ, Dowhan W (1998) The PEL1 gene (renamed PGS1) encodes the phosphatidylglycerophosphate synthase of Saccharomyces cerevisiae. J Biol Chem 273:9829–9836

    Google Scholar 

  • Chen XJ, Clark-Walker GD (1993) Mutations in MGI genes convert Kluyveromyces lactis into a petite-positive yeast. Genetics 133:517–525

    Google Scholar 

  • Chen XJ, Clark-Walker GD (1995) Specific mutations in α- and γ-subunits of F1-ATPase affect mitochondrial genome integrity in the petite-negative yeast Kluyveromyces lactis. EMBO J 14:3277–3286

    Google Scholar 

  • Chen XJ, Clark-Walker GD (1996) The mitochondrial genome integrity gene, MGI1, of Kluyveromyces lactis encodes the β-subunit of F1-ATPase. Genetics 144:1445–1454

    Google Scholar 

  • Chen XJ, Clark-Walker GD (1999) α and β subunits of F1-ATPase are required for survival of petite mutants in Saccharomyces cerevisiae. Mol Gen Genet 262:898–908

    Google Scholar 

  • Chen XJ, Clark-Walker GD (2000) The petite mutation in yeasts: 50 years on. Int Rev Cytol 194:197–238

    Google Scholar 

  • Chen XJ, Fukuhara H (1988) A gene fusion system using the aminoglycoside 3′-phosphotransferase gene of the kanamycin-resistance transposon Tn903: use in the yeast Kluyveromyces lactis and Saccharomyces cerevisiae. Gene 69:181–192

    Google Scholar 

  • Contamine V, Picard M (2000) Maintenance and integrity of the mitochondrial genome: a plethora of nuclear genes in the budding yeast. Microbiol Mol Biol Rev 64:281–315

    Google Scholar 

  • Curran SP, Leuenberger D, Oppliger W, Koehler CM (2002) The Tim9p–Tim10p complex binds to the transmembrane domains of the ADP/ATP carrier. EMBO J 21:942–953

    Google Scholar 

  • Davis AJ, Sepuri NB, Holder J, Johnson AE, Jensen RE (2000) Two intermembrane space TIM complexes interact with different domains of Tim23p during its import into mitochondria. J Cell Biol 150:1271–1282

    Google Scholar 

  • Dujon B (1981) Mitochondrial genetics and functions. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.

  • Ephrussi B (1953) Nucleocytoplasmic relations in micro-organisms: their bearing on cell heredity and differentiation. Clarendon Press, Oxford

  • Ephrussi B, Hottinguer H, Chimenes A-M (1949) Action de l′acriflavin sur les levures. I. La mutation ″petite colonie″. Ann Inst Pasteur 76:419–450

    Google Scholar 

  • Gansheroff LJ, Dollard C, Tan P, Winston F (1995) The Saccharomyces cerevisiae SPT7 gene encodes a very acidic protein important for transcription in vivo. Genetics 139:523–536

    Google Scholar 

  • Gietz RD, Schiestl RH, Willems AR, Woods RA (1995) Studies on the transformation of intact yeast cells by the LiAc/SS–DNA/PEG procedure. Yeast 11:355–360

    Google Scholar 

  • Janitor M, Subik J (1993) Molecular cloning of the PEL1 gene of Saccharomyces cerevisiae that is essential for the viability of petite mutants. Curr Genet 24:307–312

    Google Scholar 

  • Jarosch E, Tuller G, Daum G, Waldherr M, Voskova A, Schweyen RJ (1996) Mrs5p, an essential protein of the mitochondrial intermembrane space, affects protein import into yeast mitochondria. J Biol Chem 271:17219–17225

    Google Scholar 

  • Jarosch E, Rodel G, Schweyen RJ (1997) A soluble 12-kDa protein of the mitochondrial intermembrane space, Mrs11p, is essential for mitochondrial biogenesis and viability of yeast cells. Mol Gen Genet 255:157–165

    Google Scholar 

  • Kerscher O, Holder J, Srinivasan M, Leung RS, Jensen RE (1997) The Tim54p–Tim22p complex mediates insertion of proteins into the mitochondrial inner membrane. J Cell Biol 139:1663–1675

    Google Scholar 

  • Kerscher O, Sepuri NB, Jensen RE (2000) Tim18p is a new component of the Tim54p–Tim22p translocon in the mitochondrial inner membrane. Mol Biol Cell 11:103–116

    Google Scholar 

  • Koehler CM, Jarosch E, Tokatlidis K, Schmid K, Schweyen RJ, Schatz G (1998a) Import of mitochondrial carriers mediated by essential proteins of the intermembrane space. Science 279:369–373

    Google Scholar 

  • Koehler CM, Merchant S, Oppliger W, Schmid K, Jarosch E, Dolfini L, Junne T, Schatz G, Tokatlidis K (1998b) Tim9p, an essential partner subunit of Tim10p for the import of mitochondrial carrier proteins. EMBO J 17:6477–6486

    Google Scholar 

  • Koehler CM, Leuenberger D, Merchant S, Renold A, Junne T, Schatz G (1999) Human deafness dystonia syndrome is a mitochondrial disease. Proc Natl Acad Sci USA 96:2141–2146

    Google Scholar 

  • Koehler CM, Murphy MP, Bally NA, Leuenberger D, Oppliger W, Dolfini L, Junne T, Schatz G, Or E (2000) Tim18p, a new subunit of the TIM22 complex that mediates insertion of imported proteins into the yeast mitochondrial inner membrane. Mol Cell Biol 20:1187–1193

    Google Scholar 

  • Kovacova V, Irmlerova J, Kovac L (1968) Oxidative phosphorylation in yeast. IV. Combination of a nuclear mutation affecting oxidative phosphorylation with cytoplasmic mutation to respiratory deficiency. Biochim Biophys Acta 162:157–163

    Google Scholar 

  • Lefebvre-Legendre L, Vaillier J, Benabdelhak H, Velours J, Slonimski PP, Di Rago JP (2001) Identification of a nuclear gene (FMC1) required for the assembly/stability of yeast mitochondrial F1-ATPase in heat stress conditions. J Biol Chem 276:6789–6796

    Google Scholar 

  • Leuenberger D, Curran SP, Wong D, Koehler CM (2003) The role of Tim9p in the assembly of the TIM22 import complexes. Traffic 4:144–152

    Google Scholar 

  • Nishimura K, Yasumura K, Igarashi K, Harashima S, Kakinuma Y (1999a) Transcription of some PHO genes in Saccharomyces cerevisiae is regulated by Spt7p. Yeast 15:1711–1717

    Google Scholar 

  • Nishimura K, Yasumura K, Igarashi K, Kakinuma Y (1999b) Involvement of Spt7p in vacuolar polyphosphate level of Saccharomyces cerevisiae. Biochem Biophys Res Commun 257:835–838

    Google Scholar 

  • Pfanner N, Geissler A (2001) Versatility of the mitochondrial protein import machinery. Nat Rev Mol Cell Biol 2:339–349

    Google Scholar 

  • Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.

    Google Scholar 

  • Senapin S, Clark-Walker GD, Chen XJ, Seraphin B, Daugeron MC (2003) RRP20, a component of the 90S preribosome, is required for pre-18S rRNA processing in Saccharomyces cerevisiae. Nucleic Acids Res 31:2524–2533

    Google Scholar 

  • Sirrenberg C, Bauer MF, Guiard B, Neupert W, Brunner M (1996) Import of carrier proteins into the mitochondrial inner membrane mediated by Tim22. Nature 384:582–585

    Google Scholar 

  • Sirrenberg C, Endres M, Folsch H, Stuart RA, Neupert W, Brunner M (1998) Carrier protein import into mitochondria mediated by the intermembrane proteins Tim10/Mrs11 and Tim12/Mrs5. Nature 391:912–915

    Google Scholar 

  • Stribinskis V, Gao GJ, Ellis SR, Martin NC (2001) Rpm2, the protein subunit of mitochondrial RNase P in Saccharomyces cerevisiae, also has a role in the translation of mitochondrially encoded subunits of cytochrome c oxidase. Genetics 158:573–585

    Google Scholar 

  • Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

    Google Scholar 

  • Thorsness PE, White KH, Fox TD (1993) Inactivation of YME1, a member of the ftsH-SEC18-PAS1-CDC48 family of putative ATPase-encoding genes, causes increased escape of DNA from mitochondria in Saccharomyces cerevisiae. Mol Cell Biol 13:5418–5426

    Google Scholar 

  • Tranebjaerg L, Hamel BC, Gabreels FJ, Renier WO, Van Ghelue M (2000) A de novo missense mutation in a critical domain of the X-linked DDP gene causes the typical deafness-dystonia-optic atrophy syndrome. Eur J Hum Genet 8:464–467

    Google Scholar 

  • Zuo XM (2001) Characterisation of the Saccharomyces cerevisiae MGM101 gene involved in the replication of mitochondrial DNA. PhD thesis. The Australian National University, Canberra

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Acknowledgements.

We thank Drs. B. Stillman for the S. cerevisiae genomic library, C. M. Koehler for tim mutants and M. Takeda, J.-P. di Rago, and D. Nelson for plasmids containing S. cerevisiae genes. S.S. was supported by a Royal Thai Government Scholarship.

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Authors and Affiliations

  1. Molecular Genetics and Evolution Group, Research School of Biological Sciences, The Australian National University, GPO Box 475, ACT 2601, Canberra, Australia

    Saengchan Senapin & G. Desmond Clark-Walker

  2. Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390-9148, USA

    Xin Jie Chen

Authors
  1. Saengchan Senapin
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  2. Xin Jie Chen
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  3. G. Desmond Clark-Walker
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Correspondence to G. Desmond Clark-Walker.

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Communicated by M. Brunner

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Senapin, S., Chen, X.J. & Clark-Walker, G.D. Transcription of TIM9, a new factor required for the petite-positive phenotype of Saccharomyces cerevisiae, is defective in spt7 mutants. Curr Genet 44, 202–210 (2003). https://doi.org/10.1007/s00294-003-0437-9

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