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. 1993 Mar 15;90(6):2202–2206. doi: 10.1073/pnas.90.6.2202

Immunosuppressants implicate protein phosphatase regulation of K+ channels in guard cells.

S Luan 1, W Li 1, F Rusnak 1, S M Assmann 1, S L Schreiber 1
PMCID: PMC46054  PMID: 7681590

Abstract

The elevation of Ca2+ levels in the cytoplasm inactivates inward-rectifying K+ channels that play a central role in regulating the apertures of stomatal pores in higher plants. However, the mechanism for the Ca(2+)-mediated inhibition of K(+)-channel function is unknown. Using patch-clamp techniques, we show that cyclophilin-cyclosporin A and FK506-binding protein-FK506 complexes, which are highly specific inhibitors of protein phosphatase 2B (calcineurin), block Ca(2+)-induced inactivation of K+ channels in Vicia faba guard cells. A constitutively active calcineurin fragment that is Ca(2+)-independent inhibits K(+)-channel activity in the absence of Ca2+. We have also identified an endogenous Ca(2+)-dependent phosphatase activity from V. faba that is inhibited by the cyclophilin-cyclosporin A and FK506-binding protein-FK506 complexes. Our findings implicate a Ca(2+)-dependent, calcineurin-like protein phosphatase in a Ca2+ signal-transduction pathway of higher plants.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Armstrong D. L. Calcium channel regulation by calcineurin, a Ca2+-activated phosphatase in mammalian brain. Trends Neurosci. 1989 Mar;12(3):117–122. doi: 10.1016/0166-2236(89)90168-9. [DOI] [PubMed] [Google Scholar]
  2. Blatt M. R., Thiel G., Trentham D. R. Reversible inactivation of K+ channels of Vicia stomatal guard cells following the photolysis of caged inositol 1,4,5-trisphosphate. Nature. 1990 Aug 23;346(6286):766–769. doi: 10.1038/346766a0. [DOI] [PubMed] [Google Scholar]
  3. Clipstone N. A., Crabtree G. R. Identification of calcineurin as a key signalling enzyme in T-lymphocyte activation. Nature. 1992 Jun 25;357(6380):695–697. doi: 10.1038/357695a0. [DOI] [PubMed] [Google Scholar]
  4. Eckert R., Chad J. E. Inactivation of Ca channels. Prog Biophys Mol Biol. 1984;44(3):215–267. doi: 10.1016/0079-6107(84)90009-9. [DOI] [PubMed] [Google Scholar]
  5. Fairley-Grenot K., Assmann S. M. Evidence for G-Protein Regulation of Inward K+ Channel Current in Guard Cells of Fava Bean. Plant Cell. 1991 Sep;3(9):1037–1044. doi: 10.1105/tpc.3.9.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fruman D. A., Klee C. B., Bierer B. E., Burakoff S. J. Calcineurin phosphatase activity in T lymphocytes is inhibited by FK 506 and cyclosporin A. Proc Natl Acad Sci U S A. 1992 May 1;89(9):3686–3690. doi: 10.1073/pnas.89.9.3686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gasser C. S., Gunning D. A., Budelier K. A., Brown S. M. Structure and expression of cytosolic cyclophilin/peptidyl-prolyl cis-trans isomerase of higher plants and production of active tomato cyclophilin in Escherichia coli. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9519–9523. doi: 10.1073/pnas.87.24.9519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gilroy S., Read N. D., Trewavas A. J. Elevation of cytoplasmic calcium by caged calcium or caged inositol triphosphate initiates stomatal closure. Nature. 1990 Aug 23;346(6286):769–771. doi: 10.1038/346769a0. [DOI] [PubMed] [Google Scholar]
  9. Haendler B., Hofer-Warbinek R., Hofer E. Complementary DNA for human T-cell cyclophilin. EMBO J. 1987 Apr;6(4):947–950. doi: 10.1002/j.1460-2075.1987.tb04843.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Harper J. F., Sussman M. R., Schaller G. E., Putnam-Evans C., Charbonneau H., Harmon A. C. A calcium-dependent protein kinase with a regulatory domain similar to calmodulin. Science. 1991 May 17;252(5008):951–954. doi: 10.1126/science.1852075. [DOI] [PubMed] [Google Scholar]
  11. Hubbard M. J., Klee C. B. Functional domain structure of calcineurin A: mapping by limited proteolysis. Biochemistry. 1989 Feb 21;28(4):1868–1874. doi: 10.1021/bi00430a066. [DOI] [PubMed] [Google Scholar]
  12. Jin Y. J., Albers M. W., Lane W. S., Bierer B. E., Schreiber S. L., Burakoff S. J. Molecular cloning of a membrane-associated human FK506- and rapamycin-binding protein, FKBP-13. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6677–6681. doi: 10.1073/pnas.88.15.6677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kennedy M. B. Regulation of neuronal function by calcium. Trends Neurosci. 1989 Nov;12(11):417–420. doi: 10.1016/0166-2236(89)90089-1. [DOI] [PubMed] [Google Scholar]
  14. Klee C. B., Draetta G. F., Hubbard M. J. Calcineurin. Adv Enzymol Relat Areas Mol Biol. 1988;61:149–200. doi: 10.1002/9780470123072.ch4. [DOI] [PubMed] [Google Scholar]
  15. Kruse T., Tallman G., Zeiger E. Isolation of Guard Cell Protoplasts from Mechanically Prepared Epidermis of Vicia faba Leaves. Plant Physiol. 1989 Aug;90(4):1382–1386. doi: 10.1104/pp.90.4.1382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Liu J., Albers M. W., Wandless T. J., Luan S., Alberg D. G., Belshaw P. J., Cohen P., MacKintosh C., Klee C. B., Schreiber S. L. Inhibition of T cell signaling by immunophilin-ligand complexes correlates with loss of calcineurin phosphatase activity. Biochemistry. 1992 Apr 28;31(16):3896–3901. doi: 10.1021/bi00131a002. [DOI] [PubMed] [Google Scholar]
  17. Liu J., Farmer J. D., Jr, Lane W. S., Friedman J., Weissman I., Schreiber S. L. Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes. Cell. 1991 Aug 23;66(4):807–815. doi: 10.1016/0092-8674(91)90124-h. [DOI] [PubMed] [Google Scholar]
  18. MacKintosh C., Cohen P. Identification of high levels of type 1 and type 2A protein phosphatases in higher plants. Biochem J. 1989 Aug 15;262(1):335–339. doi: 10.1042/bj2620335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Miller S. G., Kennedy M. B. Regulation of brain type II Ca2+/calmodulin-dependent protein kinase by autophosphorylation: a Ca2+-triggered molecular switch. Cell. 1986 Mar 28;44(6):861–870. doi: 10.1016/0092-8674(86)90008-5. [DOI] [PubMed] [Google Scholar]
  20. O'Keefe S. J., Tamura J., Kincaid R. L., Tocci M. J., O'Neill E. A. FK-506- and CsA-sensitive activation of the interleukin-2 promoter by calcineurin. Nature. 1992 Jun 25;357(6380):692–694. doi: 10.1038/357692a0. [DOI] [PubMed] [Google Scholar]
  21. Price E. R., Zydowsky L. D., Jin M. J., Baker C. H., McKeon F. D., Walsh C. T. Human cyclophilin B: a second cyclophilin gene encodes a peptidyl-prolyl isomerase with a signal sequence. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1903–1907. doi: 10.1073/pnas.88.5.1903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Raz V., Fluhr R. Calcium Requirement for Ethylene-Dependent Responses. Plant Cell. 1992 Sep;4(9):1123–1130. doi: 10.1105/tpc.4.9.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schreiber S. L. Chemistry and biology of the immunophilins and their immunosuppressive ligands. Science. 1991 Jan 18;251(4991):283–287. doi: 10.1126/science.1702904. [DOI] [PubMed] [Google Scholar]
  24. Schreiber S. L., Crabtree G. R. The mechanism of action of cyclosporin A and FK506. Immunol Today. 1992 Apr;13(4):136–142. doi: 10.1016/0167-5699(92)90111-J. [DOI] [PubMed] [Google Scholar]
  25. Schreiber S. L., Liu J., Albers M. W., Karmacharya R., Koh E., Martin P. K., Rosen M. K., Standaert R. F., Wandless T. J. Immunophilin-ligand complexes as probes of intracellular signaling pathways. Transplant Proc. 1991 Dec;23(6):2839–2844. [PubMed] [Google Scholar]
  26. Standaert R. F., Galat A., Verdine G. L., Schreiber S. L. Molecular cloning and overexpression of the human FK506-binding protein FKBP. Nature. 1990 Aug 16;346(6285):671–674. doi: 10.1038/346671a0. [DOI] [PubMed] [Google Scholar]
  27. Swanson S. K., Born T., Zydowsky L. D., Cho H., Chang H. Y., Walsh C. T., Rusnak F. Cyclosporin-mediated inhibition of bovine calcineurin by cyclophilins A and B. Proc Natl Acad Sci U S A. 1992 May 1;89(9):3741–3745. doi: 10.1073/pnas.89.9.3741. [DOI] [PMC free article] [PubMed] [Google Scholar]

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