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Protein synthesis of Btn2 under pronounced translation repression during the process of alcoholic fermentation and wine-making in yeast

  • Applied genetics and molecular biotechnology
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Abstract

Acute high-concentration ethanol (> 9% v/v) has adverse effects on Saccharomyces cerevisiae, including the remarkable repression of bulk mRNA translation. Therefore, increased mRNA levels do not necessarily lead to an increase in the corresponding protein levels in yeast cells under severe ethanol stress. We previously identified that synthesis of Btn2 protein was efficiently induced even under the pronounced translation repression caused by acute severe ethanol stress under laboratory conditions. However, it remains to be clarified whether the translational activity is also repressed and whether the synthesis of Btn2 protein is induced during the process of alcoholic fermentation, in which the ethanol concentration increases gradually to reach high levels. In this study, we revealed that the pronounced translation repression and the translation of BTN2 are induced by high ethanol concentrations that form gradually during alcoholic fermentation using a wine yeast strain EC1118. Furthermore, we confirmed the induced expression of non-native genes driven by the BTN2 promoter during the later stage of the wine-making process. Our findings provide new information on the translation activity in yeast cells during alcoholic fermentation and suggest the utility of the BTN2 promoter for sustaining the fermentation efficiency and quality modification of alcoholic beverages.

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References

  • Alberti S (2012) Molecular mechanisms of spatial protein quality control. Prion 6:437–442

    Article  CAS  Google Scholar 

  • Ambroset C, Petit M, Brion C, Sanchez I, Delobel P, Guérin C, Chiapello H, Nicolas P, Bigey F, Dequin S, Blondin B (2011) Deciphering the molecular basis of wine yeast fermentation traits using a combined genetic and genomic approach. G3 (Bethesda) 1:263–281

    Article  Google Scholar 

  • Backhus LE, DeRisi J, Bisson LE (2001) Functional genomic analysis of a commercial wine strain of Saccharomyces cerevisiae under differing nitrogen conditions. FEMS Yeast Res 1:111–125

    Article  CAS  Google Scholar 

  • Bauer FF, Pretorius IS (2000) Yeast stress response and fermentation efficiency: how to survive the making wine—a review. S Afr J Enol Vitic 21:27–51

    CAS  Google Scholar 

  • Beltran G, Novo M, Leberre V, Sokol S, Labourdette D, Guillamon JM, Mas A, François J, Rozes N (2006) Integration of transcriptomic and metabolic analyses for understanding the global responses of low-temperature winemaking fermentations. FEMS Yeast Res 6:1167–1183

    Article  CAS  Google Scholar 

  • Duteurtre B, Bourgeois C, Chollot B (1971) Study of the assimilation of proline by brewing yeast. J Inst Brew 77:28–35

    Article  CAS  Google Scholar 

  • Espinazo-Romeu M, Cantoral JM, Matallana E, Aranda A (2008) Btn2p is involved in ethanol tolerance and biofilm formation in flor yeast. FEMS Yeast Res 8:1127–1136

    Article  CAS  Google Scholar 

  • Hashida-Okado T, Ogawa A, Endo M, Yasumoto R, Takesako K, Kato I (1996) AUR1, a novel gene conferring aureobasidin resistance on Saccharomyces cerevisiae: a study of defective morphologies in Aur1p-depleted cells. Mol Gen Genet 251:236–244

    CAS  PubMed  Google Scholar 

  • Hofmann S, Cherkasova V, Bankhead P, Bukau B, Stoecklin G (2012) Translation suppression promotes stress granule formation and cell survival in response to cold shock. Mol Biol Cell 23:3786–3800

    Article  CAS  Google Scholar 

  • Inada T, Aiba H (2005) Translation of aberrant mRNAs lacking a termination codon or with a shortened 3’-UTR is repressed after initiation in yeast. EMBO J 24:1584–1595

    Article  CAS  Google Scholar 

  • Ingledew WM, Magnus CA, Sosulski FW (1987) Influence of oxygen on proline utilization during the wine fermentation. Am J Enol Vitic 38:246–248

    CAS  Google Scholar 

  • Ishida Y, Nguyen TTM, Kitajima S, Izawa S (2016) Prioritized expression of BDH2 under bulk translational repression and its contribution to tolerance to severe vanillin stress in Saccharomyces cerevisiae. Front Microbiol 7:1059

    Article  Google Scholar 

  • Ishida Y, Nguyen TTM, Izawa S (2017) The yeast ADH7 promoter enables gene expression under pronounced translation repression caused by the combined stress of vanillin, furfural, and 5-hydroxymethylfurfural. J Biotechnol 252:65–72

    Article  CAS  Google Scholar 

  • Iwaki A, Kawai T, Yamamoto Y, Izawa S (2013) Biomass conversion inhibitors, furfural and 5-hydroxymethylfurfural, induce the formation of mRNP granules and attenuate translation activity in yeast. Appl Environ Microbiol 446:225–233

    Google Scholar 

  • Izawa S, Takemura R, Miki T, Inoue Y (2005a) Characterization of the export of bulk poly (A)+ mRNA in Saccharomyces cerevisiae during the wine-making process. Appl Environ Microbiol 71:2179–2182

    Article  CAS  Google Scholar 

  • Izawa S, Takemura R, Ikeda K, Fukuda K, Wakai Y, Inoue Y (2005b) Characterization of Rat8 localization and mRNA export in Saccharomyces cerevisiae during the brewing of Japanese sake. Appl Microbiol Biotechnol 69:86–91

    Article  CAS  Google Scholar 

  • Izawa S, Kita T, Ikeda K, Miki T, Inoue Y (2007) Formation of cytoplasmic P-bodies in sake yeast during Japanese sake brewing and wine making. Biosci Biotechnol Biochem 71:2800–2807

    Article  CAS  Google Scholar 

  • Jimenez-Marti E, del Olmo ML (2008) Addition of ammonia or amino acids to a nitrogen-depleted medium affects gene expression patterns in yeast cells during alcoholic fermentation. FEMS Yeast Res 8:245–256

    Article  CAS  Google Scholar 

  • Kama R, Robinson M, Gerst JE (2007) Btn2, a hook1 ortholog and potential batten disease-related protein, mediates late endosome-Golgi protein sorting in yeast. Mol Cell Biol 27:605–621

    Article  CAS  Google Scholar 

  • Kato K, Yamamoto Y, Izawa S (2011) Severe ethanol stress induces assembly of stress granules in Saccharomyces cerevisiae. Yeast 28:339–347

    Article  CAS  Google Scholar 

  • Lam FH, Ghaderi A, Fink GR, Stephanopoulos G (2014) Biofuels. Engineering alcohol tolerance in yeast. Science 346:71–75

    Article  CAS  Google Scholar 

  • Long D, Wilkinson KL, Taylor DK, Jiranek V (2018) Novel wine yeast for improved utilisation of proline during fermentation. Fermentation 4:10. https://doi.org/10.3390/fermentation4010010

    Article  Google Scholar 

  • Marks VD, Ho Sui SJ, Erasmus D, van der Merwe GK, Brumm J, Wasserman WW, Bryan J, van Vuuren HJ (2008) Dynamics of the yeast transcriptome during wine fermentation reveals a novel fermentation stress response. FEMS Yeast Res 8:35–52

    Article  CAS  Google Scholar 

  • Miller SB, Ho CT, Winkler J, Khokhrina M, Neuner A, Mohamed MY, Guilbride DL, Richter K, Lisby M, Schiebel E, Mogk A, Bukau B (2015) Compartment-specific aggregases direct distinct nuclear and cytoplasmic aggregate deposition. EMBO J 34:778–797

    Article  CAS  Google Scholar 

  • Nguyen TTM, Iwaki A, Izawa S (2015) The ADH7 promoter of Saccharomyces cerevisiae is vanillin-inducible and enables mRNA translation under severe vanillin stress. Front Microbiol 6:1390

    PubMed  PubMed Central  Google Scholar 

  • Nguyen TTM, Ishida Y, Kato S, Iwaki A, Izawa S (2018) The VFH1 (YLL056C) promoter is vanillin-inducible and enables mRNA translation despite pronounced translation repression caused by severe vanillin stress in Saccharomyces cerevisiae. Yeast 35:465–475

    Article  CAS  Google Scholar 

  • Novo M, Bigey F, Beyne E, Galeote V, Gavory F, Mallet S, Cambon B, Legras JL, Wincker P, Casaregola S, Dequin S (2009) Eukaryote-to-eukaryote gene transfer events revealed by the genome sequence of the wine yeast Saccharomyces cerevisiae EC1118. Proc Natl Acad Sci U S A 106:16333–16338

    Article  CAS  Google Scholar 

  • Obrig TG, Culp WJ, McKeehan WL, Hardesty B (1971) The mechanism by which cycloheximide and related glutarimide antibiotics inhibit peptide synthesis on reticulocyte ribosomes. J Biol Chem 246:174–181

    CAS  PubMed  Google Scholar 

  • Ough CS (1968) Proline content of grapes and wines. Vitis 7:321–331

    CAS  Google Scholar 

  • Ough CS, Stashak RM (1974) Further studies on proline concentration in grapes and wines. Am J Enol Vitic 25:7–12

    CAS  Google Scholar 

  • Rossignol T, Dulau L, Julien A, Blondin B (2003) Genome-wide monitoring of wine yeast gene expression during alcoholic fermentation. Yeast 20:1369–1385

    Article  CAS  Google Scholar 

  • Rossignol T, Kobi D, Jacquet-Gutfreund L, Blondin B (2009) The proteome of a wine yeast strain during fermentation, correlation with the transcriptome. J Appl Microbiol 107:47–55

    Article  CAS  Google Scholar 

  • Sopko R, Huang D, Preston N, Chua G, Papp B, Kafadar K, Snyder M, Oliver SG, Cyert M, Hughes TR, Boone C, Andrews B (2006) Mapping pathways and phenotypes by systematic gene overexpression. Mol Cell 21:319–330

    Article  CAS  Google Scholar 

  • Sørensen SPL (1907) Enzymestudien. Biochem Z 7:45–101

    Google Scholar 

  • Stines AP, Grubb J, Gockowiak H, Henschke PA, Høj PB, van Heeswijck R (2000) Proline and arginine accumulation in developing berries of Vits vinifera L in Australian vineyards: influence of vine cultivar, berry maturity and tissue type. Aust J Grape Wine Res 6:150–158

    Article  CAS  Google Scholar 

  • Takemura R, Inoue Y, Izawa S (2004) Stress response in yeast mRNA export factor: reversible changes in Rat8p localization are caused by ethanol stress not heat shock. J Cell Sci 117:4189–4197

    Article  CAS  Google Scholar 

  • Teixeira MC, Godinho CP, Cabrito TR, Mira NP, Sá-Correia I (2012) Increased expression of the yeast multidrug resistance ABC transporter Pdr18 leads to increased ethanol tolerance and ethanol production in high gravity alcoholic fermentation. Microb Cell Factories 11:98

    Article  CAS  Google Scholar 

  • ter Schure EG, van Riel NA, Verrips CT (2000) The role of ammonia metabolism in nitrogen catabolite repression in Saccharomyces cerevisiae. FEMS Microbiol Rev 24:67–83

    Article  Google Scholar 

  • Tesnière C, Brice C, Blondin B (2015) Responses of Saccharomyces cerevisiae to nitrogen starvation in wine alcoholic fermentation. Appl Microbiol Biotechnol 99:7025–7034

    Article  Google Scholar 

  • Uesono Y, Toh-E A (2002) Transient inhibition of translation initiation by osmotic stress. J Biol Chem 277:13848–13855

    Article  CAS  Google Scholar 

  • Valero E, Millán C, Ortega JM, Mauricio JC (2003) Concentration of amino acids in wine after the end of fermentation by Saccharomyces cerevisiae strains. J Sci Food Agric 83:830–835

    Article  CAS  Google Scholar 

  • Varela C, Cárdenas J, Melo F, Agosin E (2005) Quantitative analysis of wine yeast gene expression profiles under winemaking conditions. Yeast 22:369–383

    Article  CAS  Google Scholar 

  • Wu H, Zheng X, Araki Y, Sahara H, Takagi H, Shimoi H (2006) Global gene expression analysis of yeast cells during sake brewing. Appl Environ Microbiol 72:7353–7358

    Article  CAS  Google Scholar 

  • Yamamoto Y, Izawa S (2013) Adaptive response in stress granule formation and bulk translational repression upon a combined stress of mild heat shock and mild ethanol stress in yeast. Genes Cells 18:974–984

    Article  CAS  Google Scholar 

  • Yamauchi Y, Izawa S (2016) Prioritized expression of BTN2 of Saccharomyces cerevisiae under pronounced translation repression induced by severe ethanol stress. Front Microbiol 7:1319

    Article  Google Scholar 

  • Yang J, Bae JY, Lee YM, Kwon H, Moon HY, Kang HA, Yee SB, Kim W, Choi W (2011) Construction of Saccharomyces cerevisiae strains with enhanced ethanol tolerance by mutagenesis of the TATA-binding protein gene and identification of novel genes associated with ethanol tolerance. Biotechnol Bioeng 108:1776–1787

    Article  CAS  Google Scholar 

  • Zid BM, O’Shea EK (2014) Promoter sequences direct cytoplasmic localization and translation of mRNAs during starvation in yeast. Nature 514:117–121

    Article  CAS  Google Scholar 

  • Zuzuarregui A, del Olmo ML (2004) Expression of stress response genes in wine strains with different fermentative behavior. FEMS Yeast Res 4:699–710

    Article  CAS  Google Scholar 

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Acknowledgments

We are grateful to Dr. Takeo Miki and Ms. Aya Iwaki for their constructive advices.

Funding

This study was supported by the Japan Society for the Promotion of Science (grant number 26292039 to S.I.), the Nagase Science and Technology Foundation, and the Noda Institute for Scientific Research.

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Authors and Affiliations

  1. Laboratory of Microbial Technology, Graduate School of Science and Technology, Kyoto Institute of Technology, Matsugasaki, Kyoto, 606-8585, Japan

    Sae Kato, Yukina Yamauchi & Shingo Izawa

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  1. Sae Kato
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  2. Yukina Yamauchi
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  3. Shingo Izawa
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Correspondence to Shingo Izawa.

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Kato, S., Yamauchi, Y. & Izawa, S. Protein synthesis of Btn2 under pronounced translation repression during the process of alcoholic fermentation and wine-making in yeast. Appl Microbiol Biotechnol 102, 9669–9677 (2018). https://doi.org/10.1007/s00253-018-9313-x

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  • DOI: https://doi.org/10.1007/s00253-018-9313-x

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