Skip to main content
SpringerLink
Log in

TheSaccharomyces cerevisiae genes (CMP1 andCMP2) encoding calmodulin-binding proteins homologous to the catalytic subunit of mammalian protein phosphatase 2B

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Saccharomyces cerevisiae genomic clones that encode calmodulin-binding proteins were isolated by screening a λgt11 expression library using125I-labeled calmodulin as probe. Among the cloned yeast genes, we found two closely related genes (CMP1 andCMP2) that encode proteins homologous to the catalytic subunit of phosphoprotein phosphatase. The presumed CMP1 protein (62999 Da) and CMP2 protein (68496 Da) contain a 23 amino acid sequence very similar to those identified as calmodulin-binding sites in many calmodulin-regulated proteins. The yeast genes encode proteins especially homologous to the catalytic subunit of mammalian phosphoprotein phosphatase type 213 (calcineurin). The products of theCMP1 andCMP2 genes were identified by immunoblot analysis of cell extracts as proteins of 62000 and 64000 Da, respectively. Gene disruption experiments demonstrated that elimination of either or both of these genes had no effect on cell viability, indicating that these genes are not essential for normal cell growth.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bennett MK, Kennedy MB (1987) Deduced primary structure of the beta-subunit of brain type II Ca2+/calmodulin-dependent protein kinase determined by molecular cloning. Proc Natl Acad Sci USA 84:1794–1798

    Article  PubMed  CAS  Google Scholar 

  • Campbell AK (1983) Intracellular calcium: its universal role as regulator. John Wiley & Sons, Chichester

    Google Scholar 

  • Cheung WY (1980) Calmodulin plays a pivotal role in cellular regulation. Science 207:19–27

    PubMed  CAS  Google Scholar 

  • Cohen PTW (1988) Two isoforms of protein phosphatase 1 may be produced from the same gene. FEBS Lett 232:17–23

    Article  PubMed  CAS  Google Scholar 

  • Cohen P, Klee CB (eds) (1988) Molecular aspects of cellular regulation, vol 5. Elsevier, New York

    Google Scholar 

  • Cox JA, Comte M, Fitton JE, DeGrado WF (1985) The interaction of calmodulin with amphiphilic peptides. J Biol Chem 260:2527–2534

    PubMed  CAS  Google Scholar 

  • Da Cruz e Silva EF, Cohen PTW (1989) Isolation of a cDNA likely to encode a novel Ca2+-dependent/calmodulin-stimu-lated protein phosphatase. Biochim Biophys Acta 1009:293–296

    PubMed  Google Scholar 

  • Da Cruz e Silva OB, Alemany S, Campbell DG, Cohen PTW (1987) Isolation and sequence analysis of a cDNA clone encoding the entire catalytic subunit of a type-2A protein phosphatase. FEBS Lett 221:415–422

    Article  PubMed  Google Scholar 

  • Davis TD, Urdea MS, Masiarz FR, Thorner J (1986) Isolation of the yeast calmodulin gene: calmodulin is an essential protein. Cell 47:423–431

    Article  PubMed  CAS  Google Scholar 

  • Guerini D, Klee CB (1989) Cloning of human calcineurin A: evidence for two isozymes and identification of a polyproline structural domain. Proc Natl Acad Sci USA 86:9183–9187

    Article  PubMed  CAS  Google Scholar 

  • Harlow E, Lane D (1988) Antibodies: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Hiraga K, Tahara H, Taguchi N, Tsuchiya E, Fukui S, Miyakawa T (1991) Inhibition of membrane Ca2+-ATPase ofSaccharomyces cerevisiae by mating pheromone α-factor in vitro. J Gen Microbiol 137:1–4

    PubMed  CAS  Google Scholar 

  • Ito A, Hashimoto T, Hirai M, Takeda T, Shuntoh H, Kuno T, Tanaka C (1989) The complete primary structure of calcineurin A, a calmodulin binding protein homologous with protein phosphatases 1 and 2A. Biochem Biophys Res Commun 163:1492–1497

    Article  PubMed  CAS  Google Scholar 

  • Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168

    PubMed  CAS  Google Scholar 

  • Klee CB, Crouch TH, Krinks MH (1979) Calcineurin: a calcium and calmodulin-binding protein of the nervous system. Proc Natl Acad Sci USA 76:6270–6273

    Article  PubMed  CAS  Google Scholar 

  • Kronstad JW, Holly JA, MacKay VL (1987) A yeast operator overlaps an upstream activation site. Cell 50:369–377

    Article  PubMed  CAS  Google Scholar 

  • Kuno T, Takeda T, Hirai M, Ito A, Mukai H, Tanaka C (1989) Evidence for a second isoform of the catalytic subunit of calmodulin-dependent protein phosphatase (calcineurin A). Biochem Biophys Res Commun 165:1352–1358

    Article  PubMed  CAS  Google Scholar 

  • Langford CJ, Gallwitz D (1983) Evidence for an intron-contained sequence for the splicing of yeast RNA polymerase II transcripts. Cell 33:519–527

    Article  PubMed  CAS  Google Scholar 

  • Liu Y, Yamashita Y, Tsuchiya E, Miyakawa T (1990) Calmodulin-binding proteins ofSaccharomyces cerevisiae. Biochem Biophys Res Commun 166:681–686

    Article  PubMed  CAS  Google Scholar 

  • Miyakawa T, Oka Y, Tsuchiya E, Fukui S (1989)Saccharomyces cerevisiae protein kinase dependent on Ca2+ and calmodulin. J Bacteriol 171:1417–1422

    PubMed  CAS  Google Scholar 

  • Rothstein RJ (1983) One-step gene disruption in yeast. Methods Enzymol 101:202–211

    PubMed  CAS  Google Scholar 

  • Rudolph HK, Antebi A, Fink GR, Buckley CM, Dorman TE, LeVitre J, Davidow LS, Mao J, Moir DT (1989) The yeast secretory pathway is perturbed by mutations inPMR1, a member of a Ca2+ ATPase family. Cell 58:133–145

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Article  PubMed  CAS  Google Scholar 

  • Sharma RK, Taylor WA, Wang JH (1983) Use of calmodulin affinity chromatography for purification of specific calmodulin-dependent enzymes. Methods Enzymol 102:210–219

    Article  PubMed  CAS  Google Scholar 

  • Sikela JM, Hahn WE (1987) Screening an expression library with a ligand probe: isolation and sequence of a cDNA corresponding to a brain calmodulin-binding protein. Proc Natl Acad Sci USA 84:3038–3042

    Article  PubMed  CAS  Google Scholar 

  • Stanley KK (1983) Solubilization and immune-detection of β-galactosidase hybrid proteins carrying foreign antigenic determinants. Nucleic Acids Res 11:6341–6357

    Google Scholar 

  • Stewart AA, Ingebritsen TS, Manalan A, Klee CB, Cohen P (1982) Discovery of a Ca2+- and calmodulin-dependent protein phosphatase. FEBS Lett 137:80–84

    Article  PubMed  CAS  Google Scholar 

  • Takeda T, Yamamoto M (1987) Analysis andin vivo disruption of the gene coding for calmodulin inSchizosaccharomyces pombe. Proc Natl Acad Sci USA 84:3580–3584

    Article  PubMed  CAS  Google Scholar 

  • Tanaka K, Matsumoto K, Toh-e A (1988) Dual regulation of the expression of the polyubiquitin gene by cyclic AMP and heat shock in yeast. EMBO J 7:495–502

    PubMed  CAS  Google Scholar 

  • Tanaka K, Matsumoto K, Toh-e A (1989)IRA1, an inhibitory regulator of theRAS-cyclic AMP pathway inSaccharomyces cerevisiae. Mol Cell Biol 9:757–768

    PubMed  CAS  Google Scholar 

  • Yamashita I, Fukui S (1983) Molecular cloning of a glucoamylase-producing gene in the yeastSaccharomyces. Agric Biol Chem 47:2689–2692

    CAS  Google Scholar 

Download references

Author information

Author notes

  1. Sakuzo Fukui

    Present address: Department of Biotechnology, Faculty of Technology, Fukuyama University, 729-02, Fukuyama, Japan

Authors and Affiliations

  1. Department of Fermentation Technology, Faculty of Engineering, Saijo, 724, Higashi-Hiroshima, Japan

    Yusen Liu, Satoru Ishii, Masaya Tokai, Hiroko Tsutsumi, Osamu Ohki, Kazuma Tanaka, Eiko Tsuchiya, Sakuzo Fukui & Tokichi Miyakawa

  2. Center for Gene Science, Hiroshima University, Saijo, 724, Higashi-Hiroshima, Japan

    Rinji Akada

Authors
  1. Yusen Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Satoru Ishii
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Masaya Tokai
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hiroko Tsutsumi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Osamu Ohki
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Rinji Akada
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Kazuma Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Eiko Tsuchiya
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Sakuzo Fukui
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Tokichi Miyakawa
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by C.P. Hollenberg

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liu, Y., Ishii, S., Tokai, M. et al. TheSaccharomyces cerevisiae genes (CMP1 andCMP2) encoding calmodulin-binding proteins homologous to the catalytic subunit of mammalian protein phosphatase 2B. Molec. Gen. Genet. 227, 52–59 (1991). https://doi.org/10.1007/BF00260706

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00260706

Key words

Search

Navigation