Current Genetics

, Volume 28, Issue 3, pp 217–224

The S. cerevisiae nuclear gene SUV3 encoding a putative RNA helicase is necessary for the stability of mitochondrial transcripts containing multiple introns

  • Pawel Golik
  • Tomasz Szczepanek
  • Ewa Bartnik
  • Piotr P. Stepien
  • Jaga Lazowska
Original Paper

Abstract

The product of the nuclear gene SUV3 is implicated in a variety of post-transcriptional processes in yeast mitochondria. We have analysed the effect of SUV3 gene-disruption on the expression of intron-containing alleles of the mitochondrial cytb and coxl genes. We have constructed several strains with mitochondrial genomes containing different combinations of cytb and cox1 introns, and associated these genomes with the disruption of SUV3. The resulting strains were tested for their respiratory competence and spectral cytochrome content. All the strains containing only two or three introns showed normal expression of cytb and cox1, whereas the strains containing more introns were unable to express the appropriate gene. The analysis of mitochondrial RNAs by Northern hybridisation showed that the loss of respiratory competence in the strains containing more introns is due to the decrease of mRNA level with no over-accumulation of high-molecular-weight precursors. However, the transcription of the genes was not affected. These results led us to the notion that SUV3 is required for the stability of intron-containing cytb and cox1 transcripts in a cumulative way, not dependent on any particular intron.

Key words

Yeast Nucleo-mitochondrial interactions Introns RNA stability 

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References

  1. Bonitz SG, Corruzi G, Thalenfeld BE, Tzagoloff A (1980) Assembly of the mitochondrial membrane system: structure and nucleotide sequence of the gene coding for subunit 1 of yeast cytochrome oxidase. J Biol Chem 255:11927–11941Google Scholar
  2. Bousquet K, Dujardin G, Poyton RO, Slonimski PP (1990) Two group-I mitochondrial introns in the cob-box and cox1 genes require the same MRS1/PET157 nuclear gene product for splicing. Curr Genet 18:117–124Google Scholar
  3. Chang TH, Arenas J, Abelson J (1990) Identification of five putative yeast RNA helicase genes. Proc Natl Acad Sci USA 87: 1571–1575Google Scholar
  4. Claisse ML, Pere-Aubert GA, Clavilier LP, Slonimski PP (1970) Méthode d'estimation de la concentration des cytochromes dans les cellules entières de levure. Eur J Biochem 16:49–59Google Scholar
  5. Company M, Arenas J, Abelson J (1991) Requirement of the RNA helicase-like protein PRP22 for release of messenger RNA from spliceosomes. Nature 349:487–493Google Scholar
  6. Conde J, Fink G (1976) A mutant of S. cerevisiae defective for nuclear fusion. Proc Natl Acad Sci USA 73:3651–3655Google Scholar
  7. Conrad-Webb H, Perlman PS, Zhu H, Butow RA (1990) The nuclear SUV3-1 mutation affects a variety of post-transcriptional processes in yeast mitochondria. Nucleic Acids Res 18: 1369–1376Google Scholar
  8. Costanzo MC, Fox TD (1990) Control of mitochondrial gene expression in Saccharomyces cerevisiae. Annu Rev Genet 24:91–113Google Scholar
  9. Dieckmann CL, Homison G, Tzagoloff A (1984 a) 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
  10. Dieckmann CL, Koerner TJ, Tzagoloff A (1984 b) 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
  11. Dieckmann TM, Mittelmier TM (1987) Nuclearly-encoded CBP1 interacts with the 5′-end of mitochondrial cytochrome-b pre-mRNA. Curr Genet 12:391–397Google Scholar
  12. di Rago JP, Netter P, Slonimski PP (1990) Pseudo wild-type revertants from inactive apocytochrome b mutants as a tool for the analysis of the structure/function relationships of the mitochondrial ubiquinol-cytochrome c reductase of Saccharomyces cerevisiae. J Biol Chem 265:3332–3339Google Scholar
  13. Dujardin G, Pajot P, Groudinsky O, Slonimski PP (1980) Long-range control circuits within mitochondria and between nucleus and mitochondria. I. Methodology and phenomenology of suppressors. Mol Gen Genet 179:469–482Google Scholar
  14. Gorbalyenya AE, Koonin EV, Donchenko AP, Blinov VM (1988) A conserved NTP-motif in putative helicases. Nature 333:22Google Scholar
  15. Groudinsky O, Dujardin G, Slonimski PP (1981) Long-range control circuits within mitochondria and between nucleus and mitochondria. II. Genetic and biochemical analyses of suppressors which selectively alleviate the mitochondrial-intron mutations. Mol Gen Genet 184:493–503Google Scholar
  16. Groudinsky O, Bousquet I, Wallis MG, Slonimski PP, Dujardin G (1993) The NAM1/MFT2 nuclear gene product is selectively required for the stability and/or processing of mitochondrial transcripts of the atp6 and of the mosaic, cox1 and cytb genes in Saccharomyces cerevisiae. Mol Gen Genet 240:419–427Google Scholar
  17. Hensgens LAM, Bonen L, de Haan M, Horst G, Grivell LA (1983) Two introns in yeast mitochondrial COX1 gene: homology among URF-containing introns and strain-dependent variation flanking exons. Cell 32:379–389Google Scholar
  18. Hodgman TC (1988) A new superfamily of replicative proteins. Nature 333:22–23Google Scholar
  19. Iost I, Dreyfus M (1994) mRNAs can be stabilized by DEAD-box proteins. Nature 372:193–196Google Scholar
  20. Jamieson DJ, Rahe B, Pringle J, Beggs JD (1991) A suppressor of a yeast splicing mutation (prp8-1) encodes a putative ATP-dependent RNA helicase. Nature 349:715–757Google Scholar
  21. Kotylak Z, Slonimski PP (1977) Mitochondrial mutants isolated by a new screening method based upon the use of the nuclear mutation op1. In: Mitochondria W. d. Gruyter, Berlin, pp 83–89Google Scholar
  22. Kreike J, Schulze M, Pillar T, Korte A, Rodel 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
  23. Labouesse M (1990) The yeast mitochondrial leucyl-tRNA synthetase is a splicing factor for the excision of several group-I introns. Mol Gen Genet 224:209–221Google Scholar
  24. Labouresse M, Netter P, Schroeder R (1984) Molecular basis of the ‘box effect’. A maturase deficiency leading to the absence of splicing of two introns located in two split genes of yeast mitochondrial DNA. Eur J Biochem 144:85–93Google Scholar
  25. Lazowska J, Jacq C, Slonimski PP (1980) Sequence of introns and flanking exons in wild-type and box3 mutants of CYTb reveals an interlaced splicing protein coded by an intron. Cell 22:333–348Google Scholar
  26. Lazowska J, Claisse M, Gargouri A, Kotylak Z, Spyridakis A, Slonimski PP (1989) Protein encoded by the third intron of cytochrome-b gene in Saccharomyces cerevisiae is an mRNA maturase. J Mol Biol (1989) 205:275–289Google Scholar
  27. Lazowska J, Szczepanek T, Macadre C, Dokova M (1992) Two homologous mitochondrial introns from closely related Saccharomyces species differ by only a few amino-acid replacements in their open reading frames: one is mobile, the other is not. CR Acad Sci Paris 315:37–41Google Scholar
  28. Lazowska J, Meunier B, Macadre C (1994) Homing of a group-II intron in yeast mitochondrial DNA is accompanied by unidirectional co-conversion of upstream-located markers. EMBO J 13: 4963–4972Google Scholar
  29. Linder P, Slonimski PP (1989) An essential yeast protein, encoded by duplicated genes TIF1 and TIF2 and homologous to the mammalian translation initiation factor eIF-4A, can suppress a mitochondrial missense mutation. Proc Natl Acad Sci USA 86: 2286–2290Google Scholar
  30. Linder P, Lasko PF, Ashburner M, Leroy P, Nielsen PJ, Nishi K, Schnier J, Slonimski PP (1989) Birth of the D-E-A-D box. Nature 337:121–122Google Scholar
  31. 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
  32. Nobrega FG, Tzagoloff A (1980) Assembly of the mitochondrial membrane system. DNA sequence and organization of the cytochrome-b gene in Saccharomyces cerevisiae D273-10B. J Biol Chem 255, 9828–9837Google Scholar
  33. Pel HJ, Tzagoloff A, Grivell LA (1990) The identification of 18 nuclear genes required for the expression of the yeast mitochondrial gene encoding cytochrome-c oxidase subunit I. Curr Genet 21:139–146Google Scholar
  34. Schwer B, Guthrie C (1991) PRP16 is an RNA-dependent ATP-ase that interacts transiently with the spliceosome. Nature 349: 494–499Google Scholar
  35. Séraphin B, Boulet A, Simon M, Faye G (1987) Construction of a yeast strain devoid of mitochondrial introns and its use to screen nuclear genes involved in mitochondrial splicing. Proc Natl Acad Sci USA 84:6810–6814Google Scholar
  36. Séraphin B, Simon M, Faye G (1988) MSS18, a yeast nuclear gene involved in the splicing of intron aI5b of the mitochondrial cox1 transcript. EMBO J 7:1455–1464Google Scholar
  37. Séraphin B, Simon M, Boulet A, Faye G (1989) Mitochondrial splicing requires a protein from a novel helicase family. Nature 337:84–87Google Scholar
  38. Stepien PP, Margossian SP, Landsman D, Butow RA (1992) The yeast nuclear gene suv3 affecting mitochondrial post-transcriptional processes encodes a putative ATP-dependent RNA helicase. Proc Natl Acad Sci USA 89:6813–6817Google Scholar
  39. Stepien PP, Kokot L, Leski T, Bartnik E (1995) The suv3 nuclear gene product is required for the in vivo processing of the yeast mitochondrial 21-rRNA transcripts containing the r1 intron. Curr Genet 27:234–238Google Scholar
  40. Szczepanek T, Macadre C, Meunier B, Lazowska J (1994) Two homologous introns from related Saccharomyces species differ in their mobility. Gene 139:1–7Google Scholar
  41. Tian GL (1993) Recherches sur la structure et l'organisation du génome mitochondrial de la levure Saccharomyces douglasii et d'un génome mitochondrial chimérique S. douglasii et S. cerevisiae. PhD thesis, Université Pierre et Marie Curie, Paris 6, ParisGoogle Scholar
  42. Tian GL, Macadre C, Kruszewska A, Szczesniak B, Ragnini A, Grisanti P, Rinaldi T, Palleschi C, Frontali L, Slonimski PP, Lazowska J (1991) Incipient mitochondrial evolution in yeasts. I. The physical map and gene order of Saccharomyces douglasii mitochondrial DNA discloses a translocation of a segment of 15 000 base-pairs and the presence of new introns in comparison with Saccharomyces cerevisiae. J Mol Biol 218:735–746Google Scholar
  43. Tzagoloff A, Dieckman C (1990) PET genes of Saccharomyces cerevisiae. Microbial Rev 54:211–225Google Scholar
  44. Valencik ML, McEwen JE (1991) Genetic evidence that different functional domains of the PET54 gene product facilitate expression of the mitochondrial genes COX1 and COX3 in Saccharomyces cerevisiae. Mol Cell Biol 11:2399–2405Google Scholar
  45. Wenzlau JM, Saldanha RJ, Butow RA, Perlman PS (1989) A latent intron-encoded maturase is also an endonuclease needed for intron mobility. Cell 56:421–430Google Scholar
  46. Wiesenberger G, Waldherr M, Schweyen R (1992) the nuclear gene MRS2 is essential for the excision of group-II introns from yeast mitochondrial transcripts in vivo. J Biol Chem 267:6963–6969Google Scholar
  47. Zhu H, Conrad-Webb H, Liao XS, Perlman PS, Butow RA (1989) Functional expression of a yeast mitochondrial intron-encoded protein requires RNA processing at a conserved dodecamer sequence at the 3′end of the gene. Mol Cell Biol 9:1507–1512Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Pawel Golik
    • 1
    • 2
  • Tomasz Szczepanek
    • 1
  • Ewa Bartnik
    • 2
  • Piotr P. Stepien
    • 2
  • Jaga Lazowska
    • 1
  1. 1.Centre de Génétique Moléculaire du CNRSGif-sur-YvetteFrance
  2. 2.Department of GeneticsUniversity of WarsawWarsawPoland

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