Skip to main content
SpringerLink
Log in

ACR1, a gene encoding a protein related to mitochondrial carriers, is essential for acetyl-CoA synthetase activity in Saccharomyces cerevisiae

  • Original Articles
  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Abstract

The utilization of ethanol via acetate by the yeast Saccharomyces cerevisiae requires the presence of the enzyme acetyl-coenzyme A synthetase (acetyl-CoA synthetase), which catalyzes the activation of acetate to acetyl-coenzyme A (acetyl-CoA). We have isolated a mutant, termed acr1, defective for this activity by screening for mutants unable to utilize ethanol as a sole carbon source. Genetic and biochemical characterization show that, in this mutant, the structural gene for acetyl-CoA synthetase is not affected. Cloning and sequencing demonstrated that the ACR1 gene encodes a protein of 321 amino acids with a molecular mass of 35 370 Da. Computer analysis suggested that the ACR1 gene product (ACR1) is an integral membrane protein related to the family of mitochondrial carriers. The expression of the gene is induced by growing yeast cells in media containing ethanol or acetate as sole carbon sources and is repressed by glucose. ACR1 is essential for the utilization of ethanol and acetate since a mutant carrying a disruption in this gene is unable to grow on these compounds.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Adrian GS, McCammon MT, Montgomery DL, Douglas MG (1986) Sequences required for delivery and localization of the ADP/ATP translocator to the mitochondrial inner membrane. Mol Cell Biol 6:626–634

    Google Scholar 

  • Aquila H, Link TA, Klingenberg M (1985) The uncoupling protein from fat mitochondria is related to the mitochondrial ADP/ATP carrier. Analysis of sequence homologies and of folding of the protein in the membrane. EMBO J 4:2369–2376

    Google Scholar 

  • Armitt S, McCullough W, Roberts CF (1976) Analysis of acetate non-utilizing (acu) mutants in Aspergillus nidulans. J Gen Microbiol 92:263–282

    Google Scholar 

  • Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523

    Google Scholar 

  • Bolivar F, Backman K (1979) Plasmids of Escherichia coli as cloning vectors. Methods. Enzymol 68:245–250

    Google Scholar 

  • Celenza JL, Carlson M (1986) A yeast gene that is essential for release from glucose repression encodes a protein kinase. Science 233:1175–1180

    Google Scholar 

  • Celenza JL, Eng FJ, Carlson M (1989) Molecular analysis of the SNF4 gene of Saccharomyces cerevisiae: evidence for physical association of the SNF4 protein with the SNF1 protein kinase. Mol Cell Biol 9:5045–5054

    Google Scholar 

  • Ciriacy M (1975) Genetics of alcohol dehydrogenase in Saccharomyces cerevisiae. I. Isolation and genetic analysis of adh mutants. Mutat Res 29:315–326

    Google Scholar 

  • Colleaux L, Richard G-F, Thierry A, Dujon B (1992) Sequence of a segment of yeast chromosome XI identifies a new mitochondrial carrier, a new member of the G protein family, and a protein with the PAAKK motif of the H1 histones. Yeast 8:325–336

    Google Scholar 

  • Connerton IF, Fincham JRS, Sandeman RA, Hynes MJ (1990) Comparison and cross-species expression of the acetyl-CoA synthetase genes of the ascomycete fungi, Aspergillus nidulans and Neurospora crassa. Mol Microbiol 4:451–460

    Google Scholar 

  • De Virgilio C, Bürckert N, Barth G, Neuhaus J-M, Boller T, Wiemken A (1992) Cloning and disruption of a gene required for growth on acetate but not on ethanol: the acetyl-coenzyme A synthetase gene of Saccharomyces cerevisiae. Yeast 8:1043–1051

    Google Scholar 

  • Dixon GH, Kornberg HL (1959) Assay methods for key enzymes of the glyoxylate cycle. Biochem J 72:3p

    Google Scholar 

  • Erdmann R, Kunau WH (1992) A genetic approach to the biogenesis of peroxisomes in the yeast Saccharomyces cerevisiae. Cell Biochem Function 10:167–174

    Google Scholar 

  • Fernández E, Moreno F, Rodicio R (1992) The ICL1 gene from Saccharomyces cerevisiae. Eur J Biochem 204:983–990

    Google Scholar 

  • Fernández E, Fernandez M, Rodicio R (1993) Two structural genes are, encoding malate synthase isoenzymes in Saccharomyces cervisiae. FEBS Lett 320:271–275

    Google Scholar 

  • Frenkel EP, Kitchens RL (1977) Purification and properties of acetyl coenzyme A synthetase from bakers' yeast. J Biol Chem 252:504–507

    Google Scholar 

  • Gancedo JM (1992) Carbon catabolite repression in yeast. Eur J Biochem 206:297–313

    Google Scholar 

  • Gancedo C, Serrano R (1989) Energy-yielding metabolism. In: Rose AH, Harrison JS (eds) The Yeasts. Academic Press, London, pp 205–259

    Google Scholar 

  • Graf R, Baum B, Brans G (1993) YMC1, a yeast gene encoding a new putative mitochondrial carrier protein. Yeast 9:301–305

    Google Scholar 

  • Hamilton R, Watanabe CK, de Boer HA (1987) Compilation and comparison of the sequence context around the AUG start codons in Saccharomyces cerevisiae mRNAs. Annu Rev Biochem 55:913–953

    Google Scholar 

  • Hanahan D (1985) Techniques for transformation of E. coli. In: Glover DM (ed) DNA cloning: a practical approach, vol l. IRL press, Oxford, pp 109–135

    Google Scholar 

  • Hartig A, Simon MM, Schuster T, Daugherty JR, Yoo HS, Cooper TG (1992) Differentially regulated malate synthase genes participate in carbon and nitrogen metabolism of S. cerevisiae. Nucleic Acids Res 20:5677–5686

    Google Scholar 

  • Heinisch J, Hollenberg CP (1993) Yeast. In: Reed, Rehm (eds) Biotechnology, vol I. Verlagsgesellschaft. Weinheim, Germany, pp 469–514

    Google Scholar 

  • Higgins DG, Sharp PM (1988) CLUSTAL: a package for performing multiple sequence alignments on a microcomputer. Gene 73:237–244

    Google Scholar 

  • Hoffman CS, Winston F (1987) A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57:267–272

    Google Scholar 

  • Jones ME, Lippmann F (1955) Aceto-CoA-kinase. Methods Enzymol 1:585–591

    Google Scholar 

  • Klebe RJ, Harris JV, Sharp ZD, Douglas MG (1983) A general method for polyethylene glycol induced genetic transformation of bacteria and yeast. Gene 25:333–341

    Google Scholar 

  • Klein HP, Jahnke L (1979) Effects of aeration on formation and localization of the acetyl coenzyme A synthetases of Saccharomyces cerevisiae. J Bacteriol 137:179–184

    Google Scholar 

  • Kolarov J, Kolarova N, Nelson N (1990) A third ADP/ATP translocator gene in yeast. J Biol Chem 265:12711–12716

    Google Scholar 

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

    CAS  PubMed  Google Scholar 

  • Lawson JE, Douglas MG (1988) Separate genes encode functionally equivalent ADP/ATP carrier proteins in Saccharomyces cerevisiae. J Biol Chem 263:14812–14818

    Google Scholar 

  • Liao X, Butow RA (1993) RTG1 and RTG2: Two yeast genes required for a novel path of communication from mitochondria to the nucleus. Cell 72:61–71

    Google Scholar 

  • Liao X, Small WC, Srere PA, Butow RA (1991) Intramitochondrial functions regulate nonmitochondrial citrate synthase (CIT2) expression in Saccharomyces cerevisiae. Mol Cell Biol 11:38–46

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randal RJ (1951) Protein measurement with the Folin reagent. J Biol Chem 193:265–275

    Google Scholar 

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

    Google Scholar 

  • Matsuoka M, Ueda Y, Aiba S (1980) Role and control of isocitrate lyase Candida lipolitica. J Bacteriol 144:692–697

    Google Scholar 

  • Nasmyth K, Tatchell K (1980) Transposable yeast mating type loci. Cell 19:753–764

    Google Scholar 

  • Rodicio R, Zimmermann FK (1985) Cloning of maltase regulatory genes in Saccharomyces cerevisiae. Isolation of the MAL2-8 cregulatory gene. Curr Genet 9:539–545

    Google Scholar 

  • Roggenkamp R (1992) Targeting signals for protein import into peroxisomes. Cell Biochem Function 10:193–199

    Google Scholar 

  • Runswick MJ, Powell SJ, Nyren P, Walker JE (1987) Sequence of the bovine mitochondrial phosphate carrier protein: structural relationship to ADP/ATP translocase and the brown fat mitochondria uncoupling protein. EMBO J 6:1367–1373

    Google Scholar 

  • Runswick MJ, Walker JE, Bisaccia F, Jacobazzi V, Palmieri F (1990) Sequence of the bovine 2-oxoglutarate/malate carrier protein: structural relationship to other mitochondrial transport proteins. Biochemistry 29:11033–11040

    Google Scholar 

  • Sandeman RA, Hynes MJ (1989) Isolation of the facA (acetylcoenzyme A synthetase) and acuE (malate synthase) genes of Aspergillus nidulans. Mol Gen Genet 218:87–92

    Google Scholar 

  • Saraste M, Walker JE (1982) Internal sequence repeats and the path of polypeptide in mitochondrial ADP/ATP translocase. FEBS Lett 144:250–254

    Google Scholar 

  • Satyanarayana T, Chervenka CH, Klein HP (1980) Subunit specificity of the two acetyl-CoA synthetases of yeast as revealed by an immunological approach. Biochem Biophys Acta 614:601–606

    Google Scholar 

  • Seegmiller JE (1955) TPN-linked aldehyde dehydrogenase from yeast. Methods Enzymol 1:511–514

    Google Scholar 

  • Sherman F, Fink GR, Hicks JB (1986) Laboratory Course Manual for Methods in Yeast Genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Struhl K, Stinchcomb DT, Scherer S, Davis RW (1979) Highfrequency transformation of yeast: Autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci USA 76:1035–1039

    Google Scholar 

  • Subramani S (1992) Targeting of proteins into the peroxisomal matrix. J Membr Biol 125:99–106

    Google Scholar 

  • Tamaki N, Hama T (1982) Aldehyde dehydrogenase from yeasts. Methods Enzymol 89:469–473

    Google Scholar 

  • Thomas GH, Connerton IF, Fincham JRS (1988) Molecular cloning, identification and transcriptional analysis of genes involved in acetate utilization in Neurospora crassa. Mol Microbiol 2:599–606

    Google Scholar 

  • Vieira J, Messing J (1991) New pUC derived cloning vectors with different selectable markers and DNA replication origins. Gene 100:189–194

    Google Scholar 

  • Wiesenberger G, Link TA, Ashen U; Walderr M, Schweyen RJ (1991) MRS3 and MRS4, two suppressors of mtRNA splicing defect in yeast, are new members of the mitochondrial carrier family. J Mol Biol 217:23–37

    Google Scholar 

  • Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: Nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Departamento de Biologia Funcional, Area de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Oviedo, E-33006, Oviedo, Spain

    María Fernández, Ernestina Fernández & Rosaura Rodicio

Authors
  1. María Fernández
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ernestina Fernández
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rosaura Rodicio
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by C.P. Hollenberg

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fernández, M., Fernández, E. & Rodicio, R. ACR1, a gene encoding a protein related to mitochondrial carriers, is essential for acetyl-CoA synthetase activity in Saccharomyces cerevisiae . Molec. Gen. Genet. 242, 727–735 (1994). https://doi.org/10.1007/BF00283428

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00283428

Key words

Search

Navigation