Skip to main content
SpringerLink
Log in

N-Acetyltransferase Mpr1 confers ethanol tolerance on Saccharomyces cerevisiae by reducing reactive oxygen species

  • Applied Genetics and Molecular Biotechnology
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

N-Acetyltransferase Mpr1 of Saccharomyces cerevisiae can reduce intracellular oxidation levels and protect yeast cells under oxidative stress, including H2O2, heat-shock, or freeze-thaw treatment. Unlike many antioxidant enzyme genes induced in response to oxidative stress, the MPR1 gene seems to be constitutively expressed in yeast cells. Based on a recent report that ethanol toxicity is correlated with the production of reactive oxygen species (ROS), we examined here the role of Mpr1 under ethanol stress conditions. The null mutant of the MPR1 and MPR2 genes showed hypersensitivity to ethanol stress, and the expression of the MPR1 gene conferred stress tolerance. We also found that yeast cells exhibited increased ROS levels during exposure to ethanol stress, and that Mpr1 protects yeast cells from ethanol stress by reducing intracellular ROS levels. When the MPR1 gene was overexpressed in antioxidant enzyme-deficient mutants, increased resistance to H2O2 or heat shock was observed in cells lacking the CTA1, CTT1, or GPX1 gene encoding catalase A, catalase T, or glutathione peroxidase, respectively. These results suggest that Mpr1 might compensate the function of enzymes that detoxify H2O2. Hence, Mpr1 has promising potential for the breeding of novel ethanol-tolerant yeast strains.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Aguilera A, Benitez T (1985) Role of mitochondria in ethanol tolerance of Saccharomyces cerevisiae. Arch Microbiol 142:389–392

    Article  CAS  PubMed  Google Scholar 

  • Alexandre H, Rousseaux I, Charpentier C (1994) Relationship between ethanol tolerance, lipid composition and plasma membrane fluidity in Saccharomyces cerevisiae and Kloeckera apiculata. FEMS Microbiol Lett 124:17–22

    Article  CAS  PubMed  Google Scholar 

  • Buttner S, Eisenberg T, Herker E, Carmona-Gutierrez D, Kroemer G, Madeo F (2006) Why yeast cells can undergo apoptosis: death in times of peace, love, and war. J Cell Biol 175:521–525

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Casey GP, Ingledew WM (1986) Ethanol tolerance in yeasts. Crit Rev Microbiol 13:219–280

    Article  CAS  PubMed  Google Scholar 

  • Charizanis C, Juhnke H, Krems B, Entian KD (1999) The mitochondrial cytochrome c peroxidase Ccp1 of Saccharomyces cerevisiae is involved in conveying an oxidative stress signal to the transcription factor Pos9 (Skn7). Mol Gen Genet 262:437–447

    Article  CAS  PubMed  Google Scholar 

  • Chi Z, Arneborg N (1999) Relationship between lipid composition, frequency of ethanol-induced respiratory deficient mutants, and ethanol tolerance in Saccharomyces cerevisiae. J Appl Microbiol 86:1047–1052

    Article  CAS  PubMed  Google Scholar 

  • Costa V, Amorim MA, Reis E, Quintanilha A, Moradas-Ferreira P (1997) Mitochondrial superoxide dismutase is essential for ethanol tolerance of Saccharomyces cerevisiae in the post-diauxic phase. Microbiol 143:1649–1656

    Article  CAS  Google Scholar 

  • Davidson JF, Whyte B, Bissinger PH, Schiestl RH (1996) Oxidative stress is involved in heat-induced cell death in Saccharomyces cerevisiae. Proc Natl Acad Sci USA 93:5116–5121

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Du X, Takagi H (2005) N-Acetyltransferase Mpr1 conferred freeze tolerance in Saccharomyces cerevisiae by reducing reactive oxygen species. J Biochem 138:391–397

    Article  CAS  PubMed  Google Scholar 

  • Fowden L (1956) Azetidine-2-carboxylic acid: a new cyclic imino acid occurring in plants. Biochem J 64:323–332

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Furukawa K, Kitano H, Mizoguchi H, Hara S (2004) Effect of cellular inositol content on ethanol tolerance of Saccharomyces cerevisiae in sake brewing. J Biosci Bioeng 98:107–113

    Article  CAS  PubMed  Google Scholar 

  • Haliwell B, Gutteridge JM (1984) Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem J 219:1–14

    Article  Google Scholar 

  • He XJ, Fassler JS (2005) Identification of novel Yap1p and Skn7p binding sites involved in the oxidative stress response of Saccharomyces cerevisiae. Mol Microbiol 58:1454–1467

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Huh WK, Falvo JV, Gerke LC, Carroll AS, Howson RW, Weissman JS, O’Shea EK (2003) Global analysis of protein localization in budding yeast. Nature 425:686–691

    Article  CAS  PubMed  Google Scholar 

  • Ibeas JI, Jimenez J (1997) Mitochondrial DNA loss caused by ethanol in Saccharomyces flor yeasts. Appl Environ Microbiol 63:7–12

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Inoue Y, Matsuda T, Sugiyama K, Izawa S, Kimura A (1999) Genetic analysis of glutathione peroxidase in oxidative stress response of Saccharomyces cerevisiae. J Biol Chem 274:27002–27009

    Article  CAS  PubMed  Google Scholar 

  • Inoue T, Iefuji H, Fuji T, Soga H, Satoh K (2000) Cloning and characterization of a gene complementing the mutation of an ethanol-sensitive mutant of sake yeast. Biosci Biotechnol Biochem 64:229–236

    Article  CAS  PubMed  Google Scholar 

  • Jabs T (1999) Reactive oxygen intermediates as mediators of programmed cell death in plants and animals. Biochem Pharmacol 57:231–245

    Article  CAS  PubMed  Google Scholar 

  • Jiang H, English AM (2006) Phenotypic analysis of the ccp1Δ and ccp1Δ-ccp1 W191F mutant strains of Saccharomyces cerevisiae indicates that cytochrome c peroxidase functions in oxidative-stress signaling. J Inorg Biochem 100:1996–2008

    Article  CAS  PubMed  Google Scholar 

  • Kimura Y, Nakamori S, Takagi H (2002) Polymorphism of the MPR1 gene required for toxic proline analogue resistance in the Saccharomyces cerevisiae complex species. Yeast 19:1437–1445

    Article  CAS  PubMed  Google Scholar 

  • Köhrer K, Domdey H (1991) Preparation of high molecular weight RNA. In: Guthrie C, Fink GR (eds) Methods enzymology, vol. 194. Academic, San Diego, pp 398–405

    Google Scholar 

  • Kumar A, Agarwal S, Heyman JA, Matson S, Heidtman M, Piccirillo S, Umansky L, Drawid A, Jansen R, Liu Y, Cheung KH, Miller P, Gerstein M, Roeder GS, Snyder M (2002) Subcellular localization of the yeast proteome. Genes Dev 16:707–719

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lee SM, Park JW (1998) Thermosensitive phenotype of yeast mutant lacking thioredoxin peroxidase. Arch Biochem Biophys 359:99–106

    Article  CAS  PubMed  Google Scholar 

  • Lee J, Godon C, Lagniel G, Spector D, Garin J, Labarre J, Toledano MB (1999) Yap1 and Skn7 control two specialized oxidative stress response regulons in yeast. J Biol Chem 274:16040–16046

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the \( 2^{{ - \Delta \Delta Ct}} \) method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Manfredini V, Martins VD, Peralba Mdo CR, Benfato MS (2005) Adaptative response to enhanced basal oxidative damage in sod mutants from Saccharomyces cerevisiae. Mol Cell Biochem 276:175–181

    Article  CAS  PubMed  Google Scholar 

  • Mansure JJC, Panek AD, Crowe LM, Crowe JH (1994) Trehalose inhibits ethanol effects on intact yeast cells and liposomes. Biochim Biophys Acta 1191:309–316

    Article  CAS  PubMed  Google Scholar 

  • Mishra P, Prasad R (1988) Role of phospholipid head groups in ethanol tolerance of Saccharomyces cerevisiae. J Gen Microbiol 134:3205–3211

    CAS  PubMed  Google Scholar 

  • Nestelbacher R, Laun P, Vondrakova D, Pichova A, Schuller C, Breitenbach M (2000) The influence of oxygen toxicity on yeast mother cell-specific aging. Exp Gerontol 35:63–70

    Article  CAS  PubMed  Google Scholar 

  • Nomura M, Takagi H (2004) Role of the yeast novel acetyltransferase Mpr1 in oxidative stress: regulation of oxygen reactive species caused by a toxic proline catabolism intermediate. Proc Natl Acad Sci USA 101:12616–12621

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nomura M, Nakamori S, Takagi H (2003) Characterization of novel acetyltransferases found in the budding and fission yeasts that detoxifies a proline analogue, azetidine-2-carboxylic acid. J Biochem 133:67–74

    Article  CAS  PubMed  Google Scholar 

  • O’Brien KM, Dirmeier R, Engle M, Poyton RO (2004) Mitochondrial protein oxidation in yeast mutants lacking manganese-(MnSOD) or copper- and zinc-containing superoxide dismutase (CuZnSOD): evidence that MnSOD and CuZnSOD have both unique and overlapping functions in protecting mitochondrial proteins from oxidative damage. J Biol Chem 279:51817–51827

    Article  PubMed  CAS  Google Scholar 

  • Park JI, Grant CM, Attfield PA, Dawes IW (1997) The freeze-thaw stress response of the yeast Saccharomyces cerevisiae is growth phase specific and is controlled by nutritional state via the RAS-cyclic AMP signal transduction pathway. Appl Environ Microbiol 63:3818–3824

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Peterson PJ, Fowden L (1963) Different specificities of proline-activating enzymes from some plant species. Nature 200:148–151

    Article  CAS  Google Scholar 

  • Piper PW (1995) The heat shock and ethanol stress responses of yeast exhibit extensive similarity and functional overlap. FEMS Microbiol Lett 134:121–127

    Article  CAS  PubMed  Google Scholar 

  • Royall JA, Ischiropoulos H (1993) Evaluation of 2′,7′-dichlorofluorescin and dihydrorhodamine 123 as fluorescent probes for intracellular H2O2 in cultured endothelial cells. Arch Biochem Biophys 302:348–355

    Article  CAS  PubMed  Google Scholar 

  • Shichiri M, Hoshikawa C, Nakamori S, Takagi H (2001) A novel acetyltranferase found in Saccharomyces cerevisiae Σ1278b that detoxifies a proline analogue, azetidine-2-carboxylic acid. J Biol Chem 276:41998–42002

    Article  CAS  PubMed  Google Scholar 

  • Susani M, Zimniak P, Fessl F, Ruis H (1976) Localization of catalase A in vacuoles of Saccharomyces cerevisiae: evidence for the vacuolar nature of isolated “yeast peroxisomes”. Hoppe-Seyler Z Physiol Chem 357:961–970

    Article  CAS  PubMed  Google Scholar 

  • Takagi H, Shichiri M, Takemura M, Mohri M, Nakamori S (2000) Saccharomyces cerevisiae Σ1278b has novel gene of the N-acetyltransferase gene superfamily required for l-proline analogue resistance. J Bacteriol 182:4249–4256

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Takagi H, Takaoka M, Kawaguchi A, Kubo Y (2005) Effect of l-proline on sake brewing and ethanol stress in Saccharomyces cerevisiae. Appl Environ Microbiol 71:8656–8662

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tsuchiya M, Suematsu M, Suzuki H (1994) In vivo visualization of oxygen radical-dependent photoemission. Methods Enzymol 233:128–140

    Article  CAS  PubMed  Google Scholar 

  • Wong CM, Zhou Y, Ng RW, Kung HF, Jin DY (2002) Cooperation of yeast peroxiredoxins Tsa1p and Tsa2p in the cellular defense against oxidative and nitrosative stress. J Biol Chem 277:5385–5394

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgment

We wish to thank M. Shichiri and M. Nomura (Fukui Prefectural University, Fukui, Japan) for assistance in the Northern analysis and H. Shimoi (National Research Institute of Brewing, Higashihiroshima, Japan) for the discussion on this work. We also thank G. Fink and C. Kaiser (Massachusetts Institute of Technology, Cambridge, MA) for providing yeast strains L5685 and CKY263, respectively. This work was supported by a grant for the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN) and by a grant-in-aid for the Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (18380062) to H.T.

Author information

Authors and Affiliations

  1. Department of Bioscience, Fukui Prefectural University, 4-1-1 Kenjojima, Matsuoka-cho, Fukui, 910-1195, Japan

    Xiaoyi Du

  2. Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara, 630-0192, Japan

    Hiroshi Takagi

Authors
  1. Xiaoyi Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hiroshi Takagi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hiroshi Takagi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Du, X., Takagi, H. N-Acetyltransferase Mpr1 confers ethanol tolerance on Saccharomyces cerevisiae by reducing reactive oxygen species. Appl Microbiol Biotechnol 75, 1343–1351 (2007). https://doi.org/10.1007/s00253-007-0940-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-007-0940-x

Keywords

Search

Navigation