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NCA3, a nuclear gene involved in the mitochondrial expression of subunits 6 and 8 of the Fo-F1 ATP synthase of S. cerevisiae

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Abstract

Respiratory-competent nuclear mutants have been isolated which presented a cryosensitive phenotype on a non-fermentative carbon source, due to a dysfunctioning of the mitochondrial F1-Fo ATP synthase which results from a relative defect in subunits 6 and 8 of the Fo sector. Both proteins are mtDNA-encoded, but the defect is due to the simultaneous presence of a mutation in two unlinked nuclear genes (NCA2 and NCA3, for Nuclear Control of ATPase) promoting a modification of the expression of the ATP8-ATP6 co-transcript (formerly denoted AAP1-OLI2). This co-transcript matures at a unique site to give two co-transcripts of 5.2 and 4.6 kb in length: in the mutant, the 5.2-kb co-transcript was greatly lowered. NCA3 was isolated from a wild-type yeast genomic library by genetic complementation. The level of the 5.2-kb transcript, like the synthesis of subunits 6 and 8, was partly restored in the transformed strain. A 1011-nucleotide ORF was identified that encodes an hydrophilic protein of 35417 Da. Disruption of chromosomal DNA within the reading frame promoted a dramatic decrease of the 5.2-kb mRNA but did not abolish the respiratory competence of a wild-type strain. NCA3 is located on chromosome IV and produces a single 1780-b transcript.

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References

  • Ackermann SH, Tzagoloff A (1990a) ATP10, a yeast nuclear gene required for the assembly of the mitochondrial F1-Fo complex. J Biol Chem 265:9952–9959

    Google Scholar 

  • Ackermann SH, Tzagoloff A (1990b) 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 

  • Ackermann SH, Gatti DL, Gellefors P, Douglas MG, Tzagoloff A (1991) ATP13 a nuclear gene of S. cerevisiae essential for the expression of subunit 9 of the mitochondrial ATPase. FEBS Lett 278:234–238

    Google Scholar 

  • Aviv H, Leder P (1972) Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci USA 69:1408–1412

    Google Scholar 

  • Beilharz M, Cobon G, Nagley P (1982) Physiological alteration of the patterns of transcription of the oli2 region of yeast mitochondrial DNA. FEBS Lett 147:235–238

    Google Scholar 

  • Cain K, Griffiths DE (1977) Studies of energy-linked reactions. Localization of the site of action of trialkytin in yeast mitochondria. Biochem J 162:575–580

    Google Scholar 

  • Costanzo MC, Fox TD (1990) Control of mitochondrial gene expression in Saccharomyces cerevisiae. Annu Rev Genet 24:91–113

    Google Scholar 

  • Finnegan PM, Payne MJ, Keramidaris E, Lukins HB (1991) Characterization of a yeast nuclear gene, AEP2, required for accumulation of mitochondrial mRNA endocing subunit 9 of the ATP synthase. Curr Genet 20:53–61

    Google Scholar 

  • Grivell L (1989) Nucleo-mitochondrial interactions in yeast mitochondrial biogenesis. Eur J Biochem 182:477–493

    Google Scholar 

  • Groudinsky O, Bousquet I, Wallis MG, Slonimski PP, Dujardin G (1993) The NAM1/MTF2 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 S. cerevisiae. Mol Gen Genet 240:419–427

    Google Scholar 

  • Guérin M, Pélissier P (1992) A rapid method for measuring the steady state levels of mitochondrial RNA in whole mitochondria. Nucleic Acids Res 20:142

    Google Scholar 

  • Guérin B, Labbe P, Somlo M (1979) Preparation of yeast mitochondria (Saccharomyces cerevisiae) with good P/O and respiratory control ratio. Methods Enzymol 55:149–159

    Google Scholar 

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

    Google Scholar 

  • Kroczek R, Siebert E (1990) Optimisation of Northern analysis by vacuum-blotting, RNA-transfer visualisation and ultraviolet fixation. Anal Biochem 184:90–95

    Google Scholar 

  • Krusewska A, Szczesniak B (1980) Construction of isomitochondrial and isonuclear strains for recombinational analysis of mitochondrial loci in S. cerevisiae. Genet Res 35:225–229

    Google Scholar 

  • Kyte J, Doolittle E (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132

    Google Scholar 

  • Labouesse M, Dujardin G, Slonimski PP (1985) The yeast nuclear gene NAM2 is essential for the mitochondrial DNA integrity and can cure a mitochondrial RNA-maturase deficiency. Cell 41:133–143

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Manon S, Guérin M (1988) Modifications of oxidative phosphorylations in mitochondria isolated from a mutant of Saccharomyces cerevisiae. Eur J Biochem 172:205–211

    Google Scholar 

  • Manon S, Guérin M (1989) Modifications of the relative proteolipid composition in the ATP synthase of a respiratory-competent mutant of Saccharomyces cerevisiae. Biochim Biophys Acta 985:127–132

    Google Scholar 

  • Manon S, Guérin M (1993) Effect of anisotropic inhibitors of ATP synthesis in mitochondria of S. cerevisiae. Biochem Mol Biol Interact 29:375–385

    Google Scholar 

  • Manon S, Rakotomanana F, Guérin M (1988) In vivo and in vitro evidence for a proton leakage through the inner mitochondrial membrane in a mutant of S. cerevisiae. Eur J Biochem 174:399–404

    Google Scholar 

  • Marck C (1987) “DNA strider”: a “C” programm for the fast analysis of DNA and protein sequences on the Apple McIntosh family of computers. Nucleic Acids Res 16:1829–1836

    Google Scholar 

  • McIntosh M, Haynes RH (1986) Sequence and expression of the dCMP deaminase gene (DCD1) of S. cerevisiae. Mol Cell Biol 6:1711–1721

    Google Scholar 

  • McKee EE, Poyton RO (1984) Mitochondrial gene expression in S. cerevisiae. Optimal conditions for protein synthesis in isolated mitochondria. J Biol Chem 259:9320–9331

    Google Scholar 

  • Paul MF (1992) Disruption et mutagenèse du gène ATP4, codant pour une sous-unité du secteur Fo de l'ATP synthase mitochondriale de S. cerevisiae. Doctoral thesis, University of Bordeaux II, France

  • Payne MJ, Schweizer E, Lukins HB (1991) Properties of two nuclear pet mutants affecting expression of the mitochondrial olil gene of S. cerevisiae. Curr Genet 24: 126–135

    Google Scholar 

  • Pélissier PP, Camougrand NM, Manon ST, Velours GM, Guérin M (1992) Regulation by nuclear genes of the mitochondrial synthesis of subunits 6 and 8 of the ATP synthase of Saccharomyces cerevisiae. J Biol Chem 267:2467–2473

    Google Scholar 

  • Pfisterer J, Buetow DE (1981) In vitro reconstruction of the mitochondrial translation system of yeast. Proc Natl Acad Sci USA 78:4917–4921

    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

    Google Scholar 

  • Senior AE (1988) ATP synthesis by oxidative phosphorylation. Physiol Rev 68:177–231

    Google Scholar 

  • Sharp PM, Li WH (1987) The codon adaptation index a measure of directional synonymous codon usage bias, and its potential application. Nucleic Acids Res 15:1281–1295

    Google Scholar 

  • Simon M, Faye G (1984) Organization and processing of the mitochondrial oxi3/oli2 multigenic transcript in yeast. Mol Gen Genet 196:266–274

    Google Scholar 

  • Tzagoloff A (1982) Mitochondria. In: Sieheritz P (ed) Cellular organelles, Plenum Press, New York, pp 131–155

    Google Scholar 

  • Tzagoloff A, Dieckmann C (1990) Pet genes of S. cerevisiae. Microbiol Rev 54:211–225

    Google Scholar 

  • Ziaja K, Michaelis G, Lisowsky T (1993) Nuclear control of the messenger RNA expression for mitochondrial ATPase subunit 9 in a new mutant yeast. J Mol Biol 229:909–916

    Google Scholar 

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Communicated by P. P. Slonimski

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Pélissier, P., Camougrand, N., Velours, G. et al. NCA3, a nuclear gene involved in the mitochondrial expression of subunits 6 and 8 of the Fo-F1 ATP synthase of S. cerevisiae . Curr Genet 27, 409–416 (1995). https://doi.org/10.1007/BF00311209

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  • DOI: https://doi.org/10.1007/BF00311209

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