Ion Tolerance of Saccharomyces cerevisiae Lacking the Ca²⁺/CaM-Dependent Phosphatase (Calcineurin) Is Improved by Mutations in URE2 or PMA1

Corresponding author: Martha S. Cyert, Department of Biological Sciences, Stanford University, Stanford, CA 94305-5020, mcyert{at}leland.stanford.edu (E-mail).

Communicating editor: E. W. JONES

Calcineurin is a conserved, Ca²⁺/CaM-stimulated protein phosphatase required for Ca²⁺-dependent signaling in many cell types. In yeast, calcineurin is essential for growth in high concentrations of Na⁺, Li⁺, Mn²⁺, and OH^-, and for maintaining viability during prolonged treatment with mating pheromone. In contrast, the growth of calcineurin-mutant yeast is better than that of wild-type cells in the presence of high concentrations of Ca²⁺. We identified mutations that suppress multiple growth defects of calcineurin-deficient yeast (cnb1 or cna1 cna2). Mutations in URE2 suppress the sensitivity of calcineurin mutants to Na⁺, Li⁺, and Mn²⁺, and increase their survival during treatment with mating pheromone. ure2 mutations require both the transcription factor Gln3p and the Na⁺ ATPase Pmr2p to confer Na⁺ and Li⁺ tolerance. Mutations in PMA1, which encodes the yeast plasma membrane H⁺-ATPase, also suppress many growth defects of calcineurin mutants. pma1 mutants display growth phenotypes that are opposite to those of calcineurin mutants; they are resistant to Na⁺, Li⁺, and Mn²⁺, and sensitive to Ca²⁺. We also show that calcineurin mutants are sensitive to aminoglycoside antibiotics such as hygromycin B while pma1 mutants are more resistant than wild type. Furthermore, pma1 and calcineurin mutations have antagonistic effects on intracellular [Na⁺] and [Ca²⁺]. Finally, we show that yeast expressing a constitutively active allele of calcineurin display pma1-like phenotypes, and that membranes from these yeast have decreased levels of Pma1p activity. These studies further characterize the roles that URE2 and PMA1 play in regulating intracellular ion homeostasis.

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