Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

Heat shock protein HSP60 can alleviate the phenotype of mitochondrial RNA-deficient temperature-sensitive mna2 pet mutants


mna2, which belongs to the class I temperature-sensitive pet mutants that lose mitochondrial (mt)RNA at restrictive temperature, was shown by complementation and sequence determination to correspond to the gene coding for HSP60. Both mna2-1 and mna2-2, the two available alleles of mna2, have conservative single amino acid substitutions in the HSP60 gene. Valine substitutes for an alanine (position 47) in mna2-1, and an isoleucine substitutes for a valine (position 77) in mna2-2. These substitutions result in defects in respiration and in steady-state mtRNA accumulation. Wild-type hsp60 alleviates the mtRNA phenotype completely, while partially relieving the respiratory deficiency.

This is a preview of subscription content, log in to check access.


  1. Ben Asher E, Groudinsky O, Dujardin G, Altamura N, Kermorgant M, Slonimski PP (1989) Novel class of nuclear genes involved in both mRNA splicing and protein synthesis in Saccharomyces cerevisiae mitochondria. Mol Gen Genet 215:517–528

  2. Berg PE, Lewin A, Christianson T, Rabinowitz M (1979) Propagation of restriction fragments from the mitochondrial DNA of Saccharomyces cerevisiae in E. coli by means of plasmid vectors. Nucleic Acids Res 6:2133–2150

  3. Biswas TK, Getz GS (1990) Regulation of transcriptional initiation in yeast mitochondria. J Biol Chem 265:19053–19059

  4. Bonneaud N, Ozier-Kalogeropoulos O, Li G, Labouesse M, Minveille-Se'bastia L, Lacroute F (1991) A family of low and high replicative, integrative and single-stranded S. cerevisiae/E. coli shuttle vectors. Yeast 7:609–615

  5. Borst P, Grivell LA (1978) The mitochondrial genome of yeast. Cell 15:705–723

  6. Broach JR, Strathern JN, Hicks JB (1979) Transformation in yeast: development of a hybrid cloning vector and isolation of the Can1 gene. Gene 8:121–133

  7. Carle GF, Olson MV (1985) An electrophoretic karyotype for yeast. Proc Natl Acad Sci USA 82:3756–3760

  8. Cheng MY, Hartl F-U, Martin J, Pollock RA, Kalousek F, Neupert W, Hallberg EM, Hallberg RL, Horwich AL (1989) Mitochondrial heat-shock protein HSP60 is essential for assembly of proteins imported into yeast mitochondria. Nature 337:620–625

  9. Cheng MY, Hard F-U, Horwich AL (1990) The mitochondrial chaperonin hsp60 is required for its own assembly. Nature 348:455–458

  10. Cryer DR, Eccleshall R, Marmur J (1975) Isolation of yeast DNA. In: Prescott DM (ed) Methods in Cell Biology, vol 12. Academic Press, New York, pp 39–44

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

  12. 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 179:469–482

  13. Edwards JC, Levens D, Rabinowitz M (1982) Analysis of transcriptional initiation of yeast mitochondrial DNA in a homologous in vitro transcription system. Cell 31:337–346

  14. Gietz RJ, Schiestl RH (1991) Applications of high efficiency lithium acetate transformation of intact cells using single-stranded nucleic acids as carrier. Yeast 7:253–263

  15. Glab N, Wise RP, Pring DR, Jacq C, Slonimski PP (1990) Expression in Saccharomyces cerevisiae of a gene associated with cytoplasmic male sterility from maize; respiratory dysfunction and uncoupling of yeast mitochondria. Mol Gen Genet 223:24–32

  16. Grivell LA (1989) Nucleomitochondrial interactions in yeast mitochondrial biogenesis. Eur J Biochem 182:447–493

  17. Hemmingsen SM, Woolford C, Vander Vies SM, Tilly K, Dennis DT, Georgopoulos CP, Hendrix RW, Ellis RJ (1988) Homologous plant and bacterial proteins chaperone oligomeric protein assembly. Nature 333:330–334

  18. Herbert CJ, Macadre C, Becam A-M, Lazowska J, Slonimski PP (1992) The MRS1 gene of S. douglasii: co-evolution of mitochondrial introns and specific splicing proteins encoded by nuclear genes. Gene Expression 2:203–214

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

  20. Locker J, Rabinowitz M (1979) An overview of mitochondrial nucleic acids and biogenesis. Methods Enzymol 56:3–16

  21. Macino G, Tzagoloff A (1979) Assembly of mitochondrial membrane system: the DNA sequence of a mitochondrial ATPase gene in Saccharomyces cerevisiae. J Biol Chem 254:4617–4623

  22. Masters BS, Stohl LL, Clayton DA (1987) Yeast mitochondrial RNA polymerase is homologous to those encoded by bacteriophages T3 and T7. Cell 51:89–99

  23. McMullin TW, Hallberg RL (1987) A normal mitochondrial protein is selectively synthesized and accumulated during heat shock in Tetrahymena thermophilia. Mol Cell Biol 7:4414–4423

  24. McMullin TW, Hallberg RL (1988) A highly evolutionarily conserved mitochondrial protein is structurally related to the protein encoded by the Escherichia coli GroEL gene. Mol Cell Biol 8:371–380

  25. Mosse MO, Linder P, Lazowska J, Slonimski PP (1993) A comprehensive compilation of 1001 nucleotide sequences encoding proteins from the yeast Saccharomyces cerevisiae (=LIST A2). Curr Genet 23:66–91

  26. Mueller DM, Getz GS (1986a) Transcriptional regulation of the mitochondrial genome of yeast Saccharomyces cerevisiae. J Biol Chem 261:11756–11764

  27. Mueller DM, Getz GS (1986b) Steady state analysis of mitochondrial RNA after growth of yeast Saccharomyces cerevisiae under catabolite repression and derepression. J Biol Chem 261:11816–11822

  28. Mueller DM, Biswas TK, Backer J, Edwards JC, Rabinowitz M, Getz GS (1987) Temperature sensitive pet mutants in yeast Saccharomyces cerevisiae that lose mitochondrial RNA. Curr Genet 11:359–367

  29. Nasmyth KA, Reed SI (1980) Isolation of genes by complementation in yeast: molecular cloning of a cell-cycle gene. Proc Natl Acad Sci USA 77:2119–2123

  30. Reading DS, Hallberg RL, Myers AM (1989) Characterization of yeast HSP60 gene coding for a mitochondrial assembly factor. Nature 337:655–659

  31. Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labelling deoxynucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251

  32. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

  33. Slonimski PP, Perrodin G, Croft J (1968) Ethidium bromide induced mutation in yeast mitochondria: complete transformation of cells into respiratory deficient non-chromosomal ‘petites’. Biochem Biophys Res Common 30:232–239

  34. Stollet ES, Koebberl DD, Sarkar G, Sommer SS (1988) Genomic amplification with transcript sequencing. Science 239:491–494

  35. Treco DA (1989) In: Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JM, Struhl K (eds) Current Protocols in Molecular Biology, vol. 2 John Wiley and Sons, New York, pp 13.11.1–13.11.5

  36. Tzagoloff A, Macino G, Sebald W (1979) Mitochondrial genes and translation products. Annu Rev Biochem 48:419–443

  37. Vitanen PV, Lorimer GH, Seetharam R, Gupta RS, Oppenheim J, Thomas JO, Cowan NJ (1992) Mammalian mitochondrial chaperonin 60 functions as a single toroidal ring. J Biol Chem 267: 695–698

Download references

Author information

Correspondence to Arunik Sanyal.

Additional information

Communicated by D. M. Lonsdale

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Sanyal, A., Harington, A., Herbert, C.J. et al. Heat shock protein HSP60 can alleviate the phenotype of mitochondrial RNA-deficient temperature-sensitive mna2 pet mutants. Molec. Gen. Genet. 246, 56–64 (1995).

Download citation

Key words

  • Temperature-sensitive mutants
  • Heat shock protein 60
  • Conservative single amino acid substitutions
  • Mitochondrial biogenesis
  • Saccharomyces cerevisiae