Profilin is required for Ca2+ homeostasis and Ca2+-modulated bud formation in yeast

Abstract

A cls5-1 mutant of Saccharomyces cerevisiae is specifically sensitive to high concentrations of Ca2+, with elevated intracellular calcium content and altered cell morphology in the presence of 100 mM Ca2+. To reveal the mechanisms of the Ca2+-sensitive phenotype, we investigated the gene responsible and its interacting network. We demonstrated that CLS5 is identical to PFY1, encoding profilin. Involvement of profilin in the maintenance of intracellular Ca2+ homeostasis was supported by the fact that both exchangeable and non-exchangeable intracellular Ca2+ pools in the cls5-1 mutant are higher than those of the wild-type strain. Several mutations of the genes whose proteins physically interact with profilin resulted in the Ca2+-sensitive phenotype. Examination of the intracellular Ca2+ pools indicated that Bni1p, Bem1p, Rho1p, and Cla4p are also required for the maintenance of Ca2+ homeostasis. Quantitative morphological analysis revealed that the Ca2+-induced morphological changes in cls5-1 cells are similar to bem1 and cls4-1 cells. Common Ca2+-induced morphological changes were an increase in cell size and a decrease of the ratio of budded cells in the population. Since a mutation allele of cls4-1 is located in the CDC24 gene, we suggest that profilin, Bem1p, and Cdc24p are required for Ca2+-modulated bud formation. Thus, profilin is involved in Ca2+ regulation in two ways: the first is Ca2+ homeostasis by coordination with Bni1p, Bem1p, Rho1p, and Cla4p, and the second is the requirement of Ca2+ for bud formation by coordination with Bem1p and Cdc24p.