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.

This article has been cited by other articles:

				G. A. Martinez-Munoz and P. Kane Vacuolar and Plasma Membrane Proton Pumps Collaborate to Achieve Cytosolic pH Homeostasis in Yeast J. Biol. Chem., July 18, 2008; 283(29): 20309 - 20319. [Abstract] [Full Text] [PDF]

				R. A. Cramer Jr., B. Z. Perfect, N. Pinchai, S. Park, D. S. Perlin, Y. G. Asfaw, J. Heitman, J. R. Perfect, and W. J. Steinbach Calcineurin Target CrzA Regulates Conidial Germination, Hyphal Growth, and Pathogenesis of Aspergillus fumigatus Eukaryot. Cell, July 1, 2008; 7(7): 1085 - 1097. [Abstract] [Full Text] [PDF]

				A. Ruiz and J. Arino Function and Regulation of the Saccharomyces cerevisiae ENA Sodium ATPase System Eukaryot. Cell, December 1, 2007; 6(12): 2175 - 2183. [Full Text] [PDF]

				J. J. Tate and T. G. Cooper Stress-responsive Gln3 Localization in Saccharomyces cerevisiae Is Separable from and Can Overwhelm Nitrogen Source Regulation J. Biol. Chem., June 22, 2007; 282(25): 18467 - 18480. [Abstract] [Full Text] [PDF]

				S. Rodriguez-Vargas, A. Sanchez-Garcia, J. M. Martinez-Rivas, J. A. Prieto, and F. Randez-Gil Fluidization of Membrane Lipids Enhances the Tolerance of Saccharomyces cerevisiae to Freezing and Salt Stress Appl. Envir. Microbiol., January 1, 2007; 73(1): 110 - 116. [Abstract] [Full Text] [PDF]

				T. Nakayashiki, C. P. Kurtzman, H. K. Edskes, and R. B. Wickner Yeast prions [URE3] and [PSI+] are diseases PNAS, July 26, 2005; 102(30): 10575 - 10580. [Abstract] [Full Text] [PDF]

				K. A. Kafadar and M. S. Cyert Integration of Stress Responses: Modulation of Calcineurin Signaling in Saccharomyces cerevisiae by Protein Kinase A Eukaryot. Cell, October 1, 2004; 3(5): 1147 - 1153. [Abstract] [Full Text] [PDF]

				Q. Lisman, D. Urli-Stam, and J. C. M. Holthuis HOR7, a Multicopy Suppressor of the Ca2+-induced Growth Defect in Sphingolipid Mannosyltransferase-deficient Yeast J. Biol. Chem., August 27, 2004; 279(35): 36390 - 36396. [Abstract] [Full Text] [PDF]

				M. Li, S. J. Martin, V. M. Bruno, A. P. Mitchell, and D. A. Davis Candida albicans Rim13p, a Protease Required for Rim101p Processing at Acidic and Alkaline pHs Eukaryot. Cell, June 1, 2004; 3(3): 741 - 751. [Abstract] [Full Text] [PDF]

				A. M. Munson, D. H. Haydon, S. L. Love, G. L. Fell, V. R. Palanivel, and A. G. Rosenwald Yeast ARL1 encodes a regulator of K+ influx J. Cell Sci., May 1, 2004; 117(11): 2309 - 2320. [Abstract] [Full Text] [PDF]

				A. Shitamukai, D. Hirata, S. Sonobe, and T. Miyakawa Evidence for Antagonistic Regulation of Cell Growth by the Calcineurin and High Osmolarity Glycerol Pathways in Saccharomyces cerevisiae J. Biol. Chem., January 30, 2004; 279(5): 3651 - 3661. [Abstract] [Full Text] [PDF]

				T. Williams-Hart, X. Wu, and K. Tatchell Protein Phosphatase Type 1 Regulates Ion Homeostasis in Saccharomyces cerevisiae Genetics, April 1, 2002; 160(4): 1423 - 1437. [Abstract] [Full Text] [PDF]

				O. Erez and C. Kahana Screening for Modulators of Spermine Tolerance Identifies Sky1, the SR Protein Kinase of Saccharomyces cerevisiae, as a Regulator of Polyamine Transport and Ion Homeostasis Mol. Cell. Biol., January 1, 2001; 21(1): 175 - 184. [Abstract] [Full Text]

				A. Goossens, N. de la Fuente, J. Forment, R. Serrano, and F. Portillo Regulation of Yeast H+-ATPase by Protein Kinases Belonging to a Family Dedicated to Activation of Plasma Membrane Transporters Mol. Cell. Biol., October 15, 2000; 20(20): 7654 - 7661. [Abstract] [Full Text]

				F. Rusnak and P. Mertz Calcineurin: Form and Function Physiol Rev, October 1, 2000; 80(4): 1483 - 1521. [Abstract] [Full Text] [PDF]

				R. Miyamoto, R. Sugiura, S. Kamitani, T. Yada, Y. Lu, S. O. Sio, M. Asakura, A. Matsuhisa, H. Shuntoh, and T. Kuno Tol1, a Fission Yeast Phosphomonoesterase, Is an In Vivo Target of Lithium, and Its Deletion Leads to Sulfite Auxotrophy J. Bacteriol., July 1, 2000; 182(13): 3619 - 3625. [Abstract] [Full Text]

				M. J. Conboy and M. S. Cyert Luv1p/Rki1p/Tcs3p/Vps54p, a Yeast Protein That Localizes to the Late Golgi and Early Endosome, Is Required for Normal Vacuolar Morphology. Mol. Biol. Cell, July 1, 2000; 11(7): 2429 - 2443. [Abstract] [Full Text]

				M. Del Poeta, M. C. Cruz, M. E. Cardenas, J. R. Perfect, and J. Heitman Synergistic Antifungal Activities of Bafilomycin A1, Fluconazole, and the Pneumocandin MK-0991/Caspofungin Acetate (L-743,873) with Calcineurin Inhibitors FK506 and L-685,818 against Cryptococcus neoformans Antimicrob. Agents Chemother., March 1, 2000; 44(3): 739 - 746. [Abstract] [Full Text]

				B. Jiang and M. S. Cyert Identification of a Novel Region Critical for Calcineurin Function in Vivo and in Vitro J. Biol. Chem., June 25, 1999; 274(26): 18543 - 18551. [Abstract] [Full Text] [PDF]

				J. M. Mulet, M. P. Leube, S. J. Kron, G. Rios, G. R. Fink, and R. Serrano A Novel Mechanism of Ion Homeostasis and Salt Tolerance in Yeast: the Hal4 and Hal5 Protein Kinases Modulate the Trk1-Trk2 Potassium Transporter Mol. Cell. Biol., May 1, 1999; 19(5): 3328 - 3337. [Abstract] [Full Text] [PDF]

				E. Talla, R. L. de Mendonca, I. Degand, A. Goffeau, and M. Ghislain Schistosoma mansoni Ca2+-ATPase SMA2 Restores Viability to Yeast Ca2+-ATPase-deficient Strains and Functions in Calcineurin-mediated Ca2+ Tolerance J. Biol. Chem., October 23, 1998; 273(43): 27831 - 27840. [Abstract] [Full Text] [PDF]

				S. Siniossoglou, E. C. Hurt, and H. R. B. Pelham Psr1p/Psr2p, Two Plasma Membrane Phosphatases with an Essential DXDX(T/V) Motif Required for Sodium Stress Response in Yeast J. Biol. Chem., June 16, 2000; 275(25): 19352 - 19360. [Abstract] [Full Text] [PDF]

				T. M. Lamb, W. Xu, A. Diamond, and A. P. Mitchell Alkaline Response Genes of Saccharomyces cerevisiae and Their Relationship to the RIM101 Pathway J. Biol. Chem., January 12, 2001; 276(3): 1850 - 1856. [Abstract] [Full Text] [PDF]

				J. L. Crespo, K. Daicho, T. Ushimaru, and M. N. Hall The GATA Transcription Factors GLN3 and GAT1 Link TOR to Salt Stress in Saccharomyces cerevisiae J. Biol. Chem., September 7, 2001; 276(37): 34441 - 34444. [Abstract] [Full Text] [PDF]