Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

MPK1 gene is required for filamentous growth induced by isoamyl alcohol in Saccharomyces cerevisiae strains from the alcoholic fermentation


The aim of this study was to evaluate the MPK1 (SLT2) gene deletion upon filamentous growth induced by isoamyl alcohol (IAA) in two haploid industrial strains of Saccharomyces cerevisiae using oligonucleotides especially designed for a laboratory S. cerevisiae strain. The gene deletion was performed by replacing part of the open reading frames from the target gene with the KanMX gene. The recombinant strains were selected by their resistance to G418, and after deletion confirmation by polymerase chain reaction, they were cultivated in a yeast extract peptone dextrose medium + 0.5% IAA to evaluate the filamentous growth in comparison to wild strains. Mpk1 derivatives were obtained for both industrial yeasts showing the feasibility of the oligonucleotides especially designed for a laboratory strain (Σ1278b) by Martinez-Anaya et al. (In yeast, the pseudohyphal phenotype induced by isoamyl alcohol results from the operation of the morphogenesis checkpoint. J Cell Sci 116:3423–3431, 2003). The filamentation rate in these derivatives was significantly lower for both strains, as induced by IAA. This drastic reduction in the filamentation ability in the deleted strains suggests that the gene MPK1 is required for IAA-induced filamentation response. The growth curves of wild and derivative strains did not differ substantially. It is not known yet whether the switch to filamentous growth affects the fermentative characteristics of the yeast or other physiological traits. A genetically modified strain for nonfilamentous growth would be useful for these studies, and the gene MPK1 could be a target gene. The feasibility of designed oligonucleotides for this deletion in industrial yeast strains is shown.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5


  1. Ceccato-Antonini SR, Sudbery PE (2004) Filamentous growth in Saccharomyces cerevisiae. Braz J Microbiol 35:173–181

  2. Ceccato-Antonini SR, Sudbery PE, Wightman R (2005) Molecular characterization of three haploid industrial strains of Saccharomyces cerevisiae and their responses to medium stress. In: Proceedings of the “XV Simpósio Nacional de Bioprocessos”, 2–5 August. Recife, Pernambuco, Brazil

  3. Davenport KR, Sohaskey M, Kamada Y, Levin DE, Gustin MC (1995) A second osmosensing signal transduction pathway in yeast. Hypotonic shock activates the PKC1 protein kinase-regulated cell integrity pathway. J Biol Chem 270:30157–30161

  4. Gimeno CJ, Ljungdahl PO, Styles CA, Fink GR (1992) Unipolar cell divisions in the yeast Saccharomyces cerevisiae led to filamentous growth: regulation by starvation and RAS. Cell 68:1077–1090

  5. Harrison JC, Bardes ESG, Ohya Y, Lew DJ (2001) A role for the Pkc1p/Mpk1p kinase cascade in the morphogenesis checkpoint. Nat Cell Biol 3:417–420

  6. Heinisch JJ, Lorberg A, Schmitz HP, Jacoby JJ (1999) The protein kinase C-mediated MAP kinase pathway involved in the maintenance of cellular integrity in Saccharomyces cerevisiae. Mol Microbiol 32:671–680

  7. Jung U, Levin D (1999) Genome-wide analysis of gene expression regulated by the yeast cell wall integrity signaling pathway. Mol Microbiol 34:1049–1057

  8. La Valle R, Wittenberg C (2001) A role for the Swe1 checkpoint kinase during filamentous growth of Saccharomyces cerevisiae. Genetics 158:549–562

  9. Longtine MS, Mckenzie A, Demarini DJ, Shah NG, Wach A, Brachat A, Phillippsen P, Pringle JR (1998) Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Yeast 14:953–961

  10. Lorenz MC, Cutler NS, Heitman J (2000) Characterization of alcohol-induced filamentous growth in Saccharomyces cerevisiae. Mol Biol Cell 11:183–199

  11. Madden K, Sheu YJ, Baetz K, Andrews B, Snyder M (1997) SBF cell cycle regulator as a target of the yeast PKC-MAP kinase pathway. Science 275:1781–1784

  12. Martinez-Anaya C (2002) Isoamyl alcohol induces filamentous growth by decoupling budding from nuclear cycle in Saccharomyces cerevisiae. PhD thesis, University of Sheffield

  13. Martinez-Anaya C, Dickinson JR, Sudbery PE (2003) In yeast, the pseudohyphal phenotype induced by isoamyl alcohol results from the operation of the morphogenesis checkpoint. J Cell Sci 116:3423–3431

  14. Merchan S, Bernal D, Serrano R, Yenushi L (2003) Response of the Saccharomyces cerevisiae Mpk1 mitogen-activated protein kinase pathway to increases in internal turgor pressure caused by loss of Ppz protein phosphatases. Eukaryotic Cell 3:100–107

  15. Mizinuma M, Hirata D, Miyahara K, Tshuchiya E, Miyahara T (1998) Role of calcineurin and Mpk1 in regulating the onset of mitosis in budding yeast. Nature 392:303–306

  16. Shitamukai A, Hirata D, Sonobe S, Miyakawa T (2004) Evidence for antagonistic regulation of cell growth by the calcineurin and high osmolarity glycerol pathway in Saccharomyces cerevisiae. J Biol Chem 279:3651–3661

  17. White TJ, Bruns T, Lee S, Taylor J (1994) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis M, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic, New York, pp 315–321

  18. Yun DJ, Zhao Y, Pardo JM, Narasimhan ML, Damsz B, Lee H, Abad LR, D’ Urzo MP, Hasegawa PM, Bressan RA (1997) Stress proteins on the yeast cell surface determine resistance to osmotin, a plant antifungal protein. Proc Natl Acad Sci USA 94:7082–7087

Download references


The authors wish to thank Dr. Peter E. Sudbery (University of Sheffield, UK) for his helpful suggestions during the postdoc supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (no. 200678/01-9) for the second author and Dr. Claudia Martinez-Anaya for providing the oligonucleotides. The research was supported by Fundação de Amparo a Pesquisa do Estado de São Paulo (04/06889-0 and 04/06669-0 grants).

Author information

Correspondence to Sandra Regina Ceccato-Antonini.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Vancetto, G.T., Ceccato-Antonini, S.R. MPK1 gene is required for filamentous growth induced by isoamyl alcohol in Saccharomyces cerevisiae strains from the alcoholic fermentation. Appl Microbiol Biotechnol 75, 111–115 (2007).

Download citation


  • Saccharomyces cerevisiae
  • Filamentation
  • MPK1
  • Isoamyl alcohol