Current Genetics

, Volume 27, Issue 5, pp 409–416 | Cite as

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

  • P. Pélissier
  • N. Camougrand
  • G. Velours
  • M. Guèrin
Original Paper


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.

Key words

Saccharomyces cerevisiae Mitochondrial synthesis Nuclear control F1Fo-ATPase 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Ackermann SH, Tzagoloff A (1990a) ATP10, a yeast nuclear gene required for the assembly of the mitochondrial F1-Fo complex. J Biol Chem 265:9952–9959Google Scholar
  2. 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 87:4986–4990Google Scholar
  3. 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 278:234–238Google Scholar
  4. Aviv H, Leder P (1972) Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci USA 69:1408–1412Google Scholar
  5. Beilharz M, Cobon G, Nagley P (1982) Physiological alteration of the patterns of transcription of the oli2 region of yeast mitochondrial DNA. FEBS Lett 147:235–238Google Scholar
  6. Cain K, Griffiths DE (1977) Studies of energy-linked reactions. Localization of the site of action of trialkytin in yeast mitochondria. Biochem J 162:575–580Google Scholar
  7. Costanzo MC, Fox TD (1990) Control of mitochondrial gene expression in Saccharomyces cerevisiae. Annu Rev Genet 24:91–113Google Scholar
  8. 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 20:53–61Google Scholar
  9. Grivell L (1989) Nucleo-mitochondrial interactions in yeast mitochondrial biogenesis. Eur J Biochem 182:477–493Google Scholar
  10. 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 240:419–427Google Scholar
  11. Guérin M, Pélissier P (1992) A rapid method for measuring the steady state levels of mitochondrial RNA in whole mitochondria. Nucleic Acids Res 20:142Google Scholar
  12. Guérin B, Labbe P, Somlo M (1979) Preparation of yeast mitochondria (Saccharomyces cerevisiae) with good P/O and respiratory control ratio. Methods Enzymol 55:149–159Google Scholar
  13. Ito M, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168Google Scholar
  14. Kroczek R, Siebert E (1990) Optimisation of Northern analysis by vacuum-blotting, RNA-transfer visualisation and ultraviolet fixation. Anal Biochem 184:90–95Google Scholar
  15. Krusewska A, Szczesniak B (1980) Construction of isomitochondrial and isonuclear strains for recombinational analysis of mitochondrial loci in S. cerevisiae. Genet Res 35:225–229Google Scholar
  16. Kyte J, Doolittle E (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132Google Scholar
  17. 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 41:133–143Google Scholar
  18. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  19. Manon S, Guérin M (1988) Modifications of oxidative phosphorylations in mitochondria isolated from a mutant of Saccharomyces cerevisiae. Eur J Biochem 172:205–211Google Scholar
  20. 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 985:127–132Google Scholar
  21. Manon S, Guérin M (1993) Effect of anisotropic inhibitors of ATP synthesis in mitochondria of S. cerevisiae. Biochem Mol Biol Interact 29:375–385Google Scholar
  22. 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 174:399–404Google Scholar
  23. 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 16:1829–1836Google Scholar
  24. McIntosh M, Haynes RH (1986) Sequence and expression of the dCMP deaminase gene (DCD1) of S. cerevisiae. Mol Cell Biol 6:1711–1721Google Scholar
  25. McKee EE, Poyton RO (1984) Mitochondrial gene expression in S. cerevisiae. Optimal conditions for protein synthesis in isolated mitochondria. J Biol Chem 259:9320–9331Google Scholar
  26. 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, FranceGoogle Scholar
  27. 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: 126–135Google Scholar
  28. 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 267:2467–2473Google Scholar
  29. Pfisterer J, Buetow DE (1981) In vitro reconstruction of the mitochondrial translation system of yeast. Proc Natl Acad Sci USA 78:4917–4921Google Scholar
  30. Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101:202–211Google Scholar
  31. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467Google Scholar
  32. Senior AE (1988) ATP synthesis by oxidative phosphorylation. Physiol Rev 68:177–231Google Scholar
  33. Sharp PM, Li WH (1987) The codon adaptation index a measure of directional synonymous codon usage bias, and its potential application. Nucleic Acids Res 15:1281–1295Google Scholar
  34. Simon M, Faye G (1984) Organization and processing of the mitochondrial oxi3/oli2 multigenic transcript in yeast. Mol Gen Genet 196:266–274Google Scholar
  35. Tzagoloff A (1982) Mitochondria. In: Sieheritz P (ed) Cellular organelles, Plenum Press, New York, pp 131–155Google Scholar
  36. Tzagoloff A, Dieckmann C (1990) Pet genes of S. cerevisiae. Microbiol Rev 54:211–225Google Scholar
  37. 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 229:909–916Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • P. Pélissier
    • 1
  • N. Camougrand
    • 1
  • G. Velours
    • 1
  • M. Guèrin
    • 1
  1. 1.Institut de Biochimie et Génétique CellulairesUniversité de Bordeaux IIBordeauxFrance

Personalised recommendations