Dilution kinetic studies of yeast populations: in vivo aggregation of galactose utilizing enzymes and positive regulator molecules

Abstract

By use of a selective medium containing ethidium bromide, population analyses of yeast galactose long-term adaptation mutants (gal3) in the process of deadaptation in the absence of galactose have been performed. The analysis of diploid strains homozygous for the gal3 locus but heterozygous for different combinations of the other mutant galactose loci, which thus have reduced amounts of the gene products of those loci, have demonstrated that, in addition to the two permease units determined in a previous study, a cell requires one complex of the Leloir pathway enzymes and two complexes specified by the Gal4 locus to be readily induced. From the consideration of these complexes as being aggregated molecules which are diluted out as units (i.e., if such a molecule were a dimer, it would not dissociate into monomers) during cell growth, the in vivo aggregation of these enzymes and the Gal4 gene product could be studied. The data indicate that the function of the Gal4 gene product is to activate a Leloir enzyme complex. It is postulated that the gal3 phenotype is the result of such strains' inability to actively synthesize an endogenous co-inducer which allows wild-type cells to be readily induced upon exposure to galactose.