Influence of Ras function on ethanol stress response of sake yeast.
Reporter assay and Northern hybridization analysis revealed that the deletion of the RAS2 gene induced the expression of stress-responsive genes such as YAK1, CTT1, HSP12, and TSA2 in the laboratory yeast YNN27, but not in the sake yeast UT-1, suggesting that the Ras-cAMP-PKA signaling pathway does not play a very important part in the regulation of transcription of general stress-responsive genes in sake yeasts. However, these analyses showed that ethanol induces other stress-response element (STRE)-driven genes in the strain UT-1, with the exception of YAK1 which encodes a growth inhibitory protein, implying an ethanol-specific response. The good growth of the sake yeast in the presence of ethanol could be partially explained by YAK1 mRNA levels being unaffected by ethanol. Although the ras2 disruption of strain UT-1 did not potentiate ethanol tolerance, the disruptant could grow well in the presence of ethanol, and acquired ethanol tolerance, as is the case with the wild-type strain. These results suggest that specific stress responses of the sake yeast, which are different from those of the laboratory yeast, result in high ethanol tolerance and hence good growth in the presence of ethanol.
Yamaji
Kohei
K
General Research Laboratory of Kiku-Masamune Sake Brewing Co. Ltd., 1-8-6 Uozaki-nishimachi, Higashinada-ku, Kobe 658-0026, Japan.
Hara
Shodo
S
Mizoguchi
Haruhiko
H