Calcineurin acts through the CRZ1/TCN1-encoded transcription factor to regulate gene expression in yeast.

Calcineurin is a conserved Ca2+/calmodulin-dependent protein phosphatase that plays a critical role in Ca2+ signaling. We describe new components of a calcineurin-mediated response in yeast, the Ca2+-induced transcriptional activation of FKS2, which encodes a beta-1,3 glucan synthase. A 24-bp region of the FKS2 promoter was defined as sufficient to confer calcineurin-dependent transcriptional induction on a minimal promoter in response to Ca2+ and was named CDRE (for calcineurin-dependent response element). The product of CRZ1 (YNL027w) was identified as an activator of CDRE-driven transcription. Crz1p contains zinc finger motifs and binds specifically to the CDRE. Genetic analysis revealed that crz1Delta mutant cells exhibit several phenotypes similar to those of calcineurin mutants and that overexpression of CRZ1 in calcineurin mutants suppressed these phenotypes. These results suggest that Crz1p functions downstream of calcineurin to effect multiple calcineurin-dependent responses. Moreover, the calcineurin-dependent transcriptional induction of FKS2 in response to Ca2+, alpha-factor, and Na+ was found to require CRZ1. In addition, we found that the calcineurin-dependent transcriptional regulation of PMR2 and PMC1 required CRZ1. However, transcription of PMR2 and PMC1 was activated by only a subset of the treatments that activated FKS2 transcription. Thus, in response to multiple signals, calcineurin acts through the Crz1p transcription factor to differentially regulate the expression of several target genes in yeast.

[1]  D. Matheos,et al.  Tcn1p/Crz1p, a calcineurin-dependent transcription factor that differentially regulates gene expression in Saccharomyces cerevisiae. , 1997, Genes & development.

[2]  J. Mulholland,et al.  An essential role of the yeast pheromone-induced Ca2+ signal is to activate calcineurin. , 1997, Molecular biology of the cell.

[3]  E. Craig,et al.  Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. , 1996, Genetics.

[4]  P. Walter,et al.  A Novel Mechanism for Regulating Activity of a Transcription Factor That Controls the Unfolded Protein Response , 1996, Cell.

[5]  T. Davis,et al.  Ca2+-calmodulin promotes survival of pheromone-induced growth arrest by activation of calcineurin and Ca2+-calmodulin-dependent protein kinase , 1996, Molecular and cellular biology.

[6]  Thomas Fiedler,et al.  A new efficient gene disruption cassette for repeated use in budding yeast , 1996, Nucleic Acids Res..

[7]  M. Cyert,et al.  The product of HUM1, a novel yeast gene, is required for vacuolar Ca2+/H+ exchange and is related to mammalian Na+/Ca2+ exchangers , 1996, Molecular and cellular biology.

[8]  G. Fink,et al.  Calcineurin inhibits VCX1-dependent H+/Ca2+ exchange and induces Ca2+ ATPases in Saccharomyces cerevisiae , 1996, Molecular and cellular biology.

[9]  Ramón Serrano,et al.  Multiple transduction pathways regulate the sodium‐extrusion gene PMR2/ENA1 during salt stress in yeast , 1996, FEBS letters.

[10]  Y. Anraku,et al.  Yeast Cls2p/Csg2p localized on the endoplasmic reticulum membrane regulates a non‐exchangeable intracellular Ca2+ pool cooperatively with calcineurin , 1996, FEBS letters.

[11]  N. Morin,et al.  Differential expression and function of two homologous subunits of yeast 1,3-beta-D-glucan synthase , 1995, Molecular and cellular biology.

[12]  A. Futcher,et al.  Use of polymerase chain reaction epitope tagging for protein tagging in Saccharomyces cerevisiae , 1995, Yeast.

[13]  F. Maxfield,et al.  Ca2+- and calcineurin-dependent recycling of an integrin to the front of migrating neutrophils , 1995, Nature.

[14]  D. Hirata,et al.  Protein phosphatase 2B of Saccharomyces cerevisiae is required for tolerance to manganese, in blocking the entry of ions into the cells. , 1995, European journal of biochemistry.

[15]  M. Cyert,et al.  Calcineurin, the Ca2+/calmodulin-dependent protein phosphatase, is essential in yeast mutants with cell integrity defects and in mutants that lack a functional vacuolar H(+)-ATPase , 1995, Molecular and cellular biology.

[16]  CE Jahr,et al.  Synaptic desensitization of NMDA receptors by calcineurin , 1995, Science.

[17]  G. Fink,et al.  Mutations in PMR1 suppress oxidative damage in yeast cells lacking superoxide dismutase , 1995, Molecular and cellular biology.

[18]  D. Clapham,et al.  Calcium signaling , 1995, Cell.

[19]  M. Perutz Polar zippers: their role in human disease. , 1995, Protein science : a publication of the Protein Society.

[20]  M. Mclaughlin,et al.  The yeast FKS1 gene encodes a novel membrane protein, mutations in which confer FK506 and cyclosporin A hypersensitivity and calcineurin-dependent growth. , 1994, Gene.

[21]  N. Morin,et al.  The Saccharomyces cerevisiae FKS1 (ETG1) gene encodes an integral membrane protein which is a subunit of 1,3-beta-D-glucan synthase. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[22]  I. Módy,et al.  Regulation of NMDA channel function by endogenous Ca2+-dependent phosphatase , 1994, Nature.

[23]  F. Rubio,et al.  The protein phosphatase calcineurin is essential for NaCl tolerance of Saccharomyces cerevisiae. , 1994, The Journal of biological chemistry.

[24]  G. Fink,et al.  Calcineurin-dependent growth control in Saccharomyces cerevisiae mutants lacking PMC1, a homolog of plasma membrane Ca2+ ATPases , 1994, The Journal of cell biology.

[25]  I. Herskowitz,et al.  Isolation of ORC6, a component of the yeast origin recognition complex by a one-hybrid system. , 1993, Science.

[26]  G. Hannon,et al.  Isolation of the Rb-related p130 through its interaction with CDK2 and cyclins. , 1993, Genes & development.

[27]  Y. Liu,et al.  Protein phosphatase type 2B (calcineurin)‐mediated, FK506‐sensitive regulation of intracellular ions in yeast is an important determinant for adaptation to high salt stress conditions. , 1993, The EMBO journal.

[28]  T. Curran,et al.  The T-cell transcription factor NFATp is a substrate for calcineurin and interacts with Fos and Jun , 1993, Nature.

[29]  G. Crabtree,et al.  Characterization of the nuclear and cytoplasmic components of the lymphoid-specific nuclear factor of activated T cells (NF-AT) complex. , 1993, The Journal of biological chemistry.

[30]  C. Klee,et al.  Inhibition of neutrophil chemokinesis on vitronectin by inhibitors of calcineurin. , 1992, Science.

[31]  P. Greengard,et al.  Calcineurin mediates alpha-adrenergic stimulation of Na+,K(+)-ATPase activity in renal tubule cells. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[32]  J R Desjarlais,et al.  Toward rules relating zinc finger protein sequences and DNA binding site preferences. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[33]  M. Cyert,et al.  Regulatory subunit (CNB1 gene product) of yeast Ca2+/calmodulin-dependent phosphoprotein phosphatases is required for adaptation to pheromone , 1992, Molecular and cellular biology.

[34]  G. Crabtree,et al.  Identification of calcineurin as a key signalling enzyme in T-lymphocyte activation , 1992, Nature.

[35]  E. A. O'neill,et al.  FK-506- and CsA-sensitive activation of the interleukin-2 promoter by calcineurin , 1992, Nature.

[36]  T. Kuno,et al.  cDNA cloning of a calcineurin B homolog in Saccharomyces cerevisiae. , 1991, Biochemical and biophysical research communications.

[37]  A. Rodríguez-Navarro,et al.  A novel P‐type ATPase from yeast involved in sodium transport , 1991, FEBS letters.

[38]  Stuart L. Schreiber,et al.  Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes , 1991, Cell.

[39]  M. Cyert,et al.  Yeast has homologs (CNA1 and CNA2 gene products) of mammalian calcineurin, a calmodulin-regulated phosphoprotein phosphatase. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[40]  Dan S. Prestridge,et al.  SIGNAL SCAN: a computer program that scans DNA sequences for eukaryotic transcriptional elements , 1991, Comput. Appl. Biosci..

[41]  G. Roeder,et al.  Meiotic gene conversion and crossing over: Their relationship to each other and to chromosome synapsis and segregation , 1990, Cell.

[42]  Y. Anraku,et al.  Essential role for induced Ca2+ influx followed by [Ca2+]i rise in maintaining viability of yeast cells late in the mating pheromone response pathway. A study of [Ca2+]i in single Saccharomyces cerevisiae cells with imaging of fura-2. , 1990, The Journal of biological chemistry.

[43]  G. Fink,et al.  The yeast secretory pathway is perturbed by mutations in PMR1, a member of a Ca2+ ATPase family , 1989, Cell.

[44]  R. Sikorski,et al.  A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. , 1989, Genetics.

[45]  C. Klee,et al.  Functional domain structure of calcineurin A: mapping by limited proteolysis. , 1989, Biochemistry.

[46]  R. Evans,et al.  Zinc fingers: Gilt by association , 1988, Cell.

[47]  A. Myers,et al.  Yeast/E. coli shuttle vectors with multiple unique restriction sites , 1986, Yeast.

[48]  G K Lewis,et al.  Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product , 1985, Molecular and cellular biology.

[49]  Andrew R. Cherenson,et al.  The structure of an antigenic determinant in a protein , 1984, Cell.

[50]  L. Guarente,et al.  Heme regulates transcription of the CYC1 gene of S. cerevisiae via an upstream activation site , 1983, Cell.

[51]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[52]  Jeffrey H. Miller Experiments in molecular genetics , 1972 .