Abstract
In Saccharomyces cerevisiae, inactivation of the nuclear gene YME1 causes several phenotypes associated with impairment of mitochondrial function. In addition to deficiencies in mitochondrial compartment integrity and respiratory growth, yme1 mutants grow extremely slowly in the absence of mitochondrial DNA. We have identified two genetic loci that, when mutated, act as dominant suppressors of the slow-growth phenotype of yme1 strains lacking mitochondrial DNA. These mutations only suppressed the slow-growth phenotype of yme1 strains lacking mitochondrial DNA and had no effect on other phenotypes associated with yme1 mutations. One allele of one linkage group had a collateral respiratory deficient phenotype that allowed the isolation of the wild-type gene. This suppressing mutation was in ATP3, a gene that encodes the gamma subunit of the mitochondrial ATP synthase. Recovery of two of the suppressing ATP3 alleles and subsequent sequence analysis placed the suppressing mutations at strictly conserved residues near the C terminus of Atp3p. Deletion of the ATP3 genomic locus resulted in an inability to utilize nonfermentable carbon sources. atp3 deletion strains lacking mitochondrial DNA grew slowly on glucose media but were not as compromised for growth as yme1 yeast lacking mitochondrial DNA.
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- Baker K. P., Schatz G. Mitochondrial proteins essential for viability mediate protein import into yeast mitochondria. Nature. 1991 Jan 17;349(6306):205–208. doi: 10.1038/349205a0. [DOI] [PubMed] [Google Scholar]
- Bilofsky H. S., Burks C. The GenBank genetic sequence data bank. Nucleic Acids Res. 1988 Mar 11;16(5):1861–1863. doi: 10.1093/nar/16.5.1861. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Campbell C. L., Tanaka N., White K. H., Thorsness P. E. Mitochondrial morphological and functional defects in yeast caused by yme1 are suppressed by mutation of a 26S protease subunit homologue. Mol Biol Cell. 1994 Aug;5(8):899–905. doi: 10.1091/mbc.5.8.899. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen X. J., Clark-Walker G. D. Mutations in MGI genes convert Kluyveromyces lactis into a petite-positive yeast. Genetics. 1993 Mar;133(3):517–525. doi: 10.1093/genetics/133.3.517. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chow T. Y., Kunz B. A. Evidence that an endo-exonuclease controlled by the NUC2 gene functions in the induction of 'petite' mutations in Saccharomyces cerevisiae. Curr Genet. 1991 Jul;20(1-2):39–44. doi: 10.1007/BF00312763. [DOI] [PubMed] [Google Scholar]
- Clarkson G. H., Poyton R. O. A role for membrane potential in the biogenesis of cytochrome c oxidase subunit II, a mitochondrial gene product. J Biol Chem. 1989 Jun 15;264(17):10114–10118. [PubMed] [Google Scholar]
- Desjardins P., Frost E., Morais R. Ethidium bromide-induced loss of mitochondrial DNA from primary chicken embryo fibroblasts. Mol Cell Biol. 1985 May;5(5):1163–1169. doi: 10.1128/mcb.5.5.1163. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haffter P., Fox T. D. Nuclear mutations in the petite-negative yeast Schizosaccharomyces pombe allow growth of cells lacking mitochondrial DNA. Genetics. 1992 Jun;131(2):255–260. doi: 10.1093/genetics/131.2.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
- King M. P., Attardi G. Human cells lacking mtDNA: repopulation with exogenous mitochondria by complementation. Science. 1989 Oct 27;246(4929):500–503. doi: 10.1126/science.2814477. [DOI] [PubMed] [Google Scholar]
- Kolarov J., Kolarova N., Nelson N. A third ADP/ATP translocator gene in yeast. J Biol Chem. 1990 Jul 25;265(21):12711–12716. [PubMed] [Google Scholar]
- Kovácová V., Irmlerová J., Kovác L. Oxidative phosphorylatiion in yeast. IV. Combination of a nuclear mutation affecting oxidative phosphorylation with cytoplasmic mutation to respiratory deficiency. Biochim Biophys Acta. 1968 Aug 20;162(2):157–163. doi: 10.1016/0005-2728(68)90097-2. [DOI] [PubMed] [Google Scholar]
- McConnell S. J., Stewart L. C., Talin A., Yaffe M. P. Temperature-sensitive yeast mutants defective in mitochondrial inheritance. J Cell Biol. 1990 Sep;111(3):967–976. doi: 10.1083/jcb.111.3.967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nakamoto R. K., Maeda M., Futai M. The gamma subunit of the Escherichia coli ATP synthase. Mutations in the carboxyl-terminal region restore energy coupling to the amino-terminal mutant gamma Met-23-->Lys. J Biol Chem. 1993 Jan 15;268(2):867–872. [PubMed] [Google Scholar]
- Pajic A., Tauer R., Feldmann H., Neupert W., Langer T. Yta10p is required for the ATP-dependent degradation of polypeptides in the inner membrane of mitochondria. FEBS Lett. 1994 Oct 17;353(2):201–206. doi: 10.1016/0014-5793(94)01046-3. [DOI] [PubMed] [Google Scholar]
- Paul M. F., Ackerman S., Yue J., Arselin G., Velours J., Tzagolof A., Ackermann S [corrected to Ackerman S. ]. Cloning of the yeast ATP3 gene coding for the gamma-subunit of F1 and characterization of atp3 mutants. J Biol Chem. 1994 Oct 21;269(42):26158–26164. [PubMed] [Google Scholar]
- Pedersen P. L., Amzel L. M. ATP synthases. Structure, reaction center, mechanism, and regulation of one of nature's most unique machines. J Biol Chem. 1993 May 15;268(14):9937–9940. [PubMed] [Google Scholar]
- Rose M. D., Novick P., Thomas J. H., Botstein D., Fink G. R. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector. Gene. 1987;60(2-3):237–243. doi: 10.1016/0378-1119(87)90232-0. [DOI] [PubMed] [Google Scholar]
- Saraste M., Gay N. J., Eberle A., Runswick M. J., Walker J. E. The atp operon: nucleotide sequence of the genes for the gamma, beta, and epsilon subunits of Escherichia coli ATP synthase. Nucleic Acids Res. 1981 Oct 24;9(20):5287–5296. doi: 10.1093/nar/9.20.5287. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zanotti F., Guerrieri F., Capozza G., Fiermonte M., Berden J., Papa S. Role of F0 and F1 subunits in the gating and coupling function of mitochondrial H(+)-ATP synthase. The effect of dithiol reagents. Eur J Biochem. 1992 Aug 15;208(1):9–16. doi: 10.1111/j.1432-1033.1992.tb17153.x. [DOI] [PubMed] [Google Scholar]