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 (Sigma1278b) 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.
Vancetto
Guilherme Tadeu
GT
Departamento de Tecnologia Agroindustrial e Sócio-Economia Rural, Centro de Ciências Agrárias, Universidade Federal de São Carlos-Campus de Araras, Araras, São Paulo, Brazil.
Ceccato-Antonini
Sandra Regina
SR
D005285:Fermentation; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D005818:Genetic Engineering; D007218:Industrial Microbiology; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics* / Q000378:metabolism*; D000439:Pentanols / Q000378:metabolism / Q000494:pharmacology*; D011494:Protein Kinases / Q000378:metabolism; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000254:growth & development*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*
D000439:Pentanols; D029701:Saccharomyces cerevisiae Proteins; C029683:isopentyl alcohol; D011494:Protein Kinases; D020928:Mitogen-Activated Protein Kinases; C072724:SLT2 protein, S cerevisiae