The mechanism of inducer formation in gal3 mutants of the yeast galactose system is independent of normal galactose metabolism and mitochondrial respiratory function.
Saccharomyces cerevisiae cells defective in GAL3 function exhibit either one of two phenotypes. The gal3 mutation in an otherwise normal cell causes a 2-5-day delay in the galactose triggered induction of GAL/MEL gene transcription. This long term adaptation (LTA) phenotype has been ascribed to inefficient inducer formation. The gal3 mutation causes a noninducible phenotype for GAL/MEL transcription if cells are defective in Leloir pathway function, in glycolysis or in respiratory function. It was recently shown that multiple copies of the intact GAL1 gene partially suppress the LTA phenotype of gal3 cells. Here we report that constitutively expressed GAL1 restored gal3 mutants to the rapidly inducible phenotype characteristic of wild-type cells and conferred rapid inducibility to gal3 gal10, gal3 gal7 or gal3 rho- strains that are normally noninducible. As shown by immunoblot analysis, the GAL1-mediated induction exhibits phosphorylation of the GAL4 protein, suggesting a mechanism similar to GAL3-mediated induction. Altogether our results indicate that the deciding factor in the inducibility of the GAL/MEL genes in gal3 strains is the Gal3p-like activity of Gal1p. Based on the above we conclude that inducer formation does not require normal metabolism of galactose nor does it require mitochondrial respiratory function. These conclusions vitiate previous explanations for gal3 associated long-term adaptation and noninducible phenotypes.
Bhat
P J
PJ
Department of Biological Chemistry, Milton S. Hershey Medical Center, Pennsylvania State University, Hershey 17033.
Hopper
J E
JE
D003001:Cloning, Molecular; D005690:Galactose / Q000235:genetics* / Q000378:metabolism; D005696:Galactosidases / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D007700:Kinetics; D008928:Mitochondria / Q000378:metabolism*; D009154:Mutation; D010766:Phosphorylation; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*
D005696:Galactosidases; D005690:Galactose