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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1994 Jul 5;91(14):6310–6314. doi: 10.1073/pnas.91.14.6310

A membrane-delimited pathway of G-protein regulation of the guard-cell inward K+ channel.

W H Wu 1, S M Assmann 1
PMCID: PMC44191  PMID: 8022777

Abstract

GTP-binding protein (G-protein) regulation of inward rectifying K+ channels in the plasma membrane of Vicia (Vicia faba L.) guard cells has previously been demonstrated at the whole-cell level. However, whether a cytosolic signal transduction chain is required for G-protein regulation of K+ channels in Vicia guard cells, or in any plant cell type, remains unknown. In the present study, we assayed effects of several G-protein regulators on inward K+ channels in isolated inside-out membrane patches from Vicia guard cell protoplasts. Guanosine 5'-[gamma-thio]triphosphate, a nonhydrolyzable GTP analog that locks G proteins into their activated state, decreased the open state probability (Po) of single inward K+ channels. This decrease in Po was accompanied by an increase in one of the closed time constants of the K+ channel. Guanosine 5'-[beta-thio]diphosphate, a GDP analog that locks G proteins into their inactivated state, slightly increased the Po of the inward K+ channel and shortened the closed time constants. Pertussis toxin and cholera toxin, which ADP-ribosylate G proteins at different sites, decreased the Po of the inward K+ channel. Our data indicate that G proteins can act via a membrane-delimited pathway to regulate inward K+ channels in the guard-cell plasma membrane.

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Selected References

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  1. Assmann S. M. Signal transduction in guard cells. Annu Rev Cell Biol. 1993;9:345–375. doi: 10.1146/annurev.cb.09.110193.002021. [DOI] [PubMed] [Google Scholar]
  2. Barry P. H., Lynch J. W. Liquid junction potentials and small cell effects in patch-clamp analysis. J Membr Biol. 1991 Apr;121(2):101–117. doi: 10.1007/BF01870526. [DOI] [PubMed] [Google Scholar]
  3. Blum W., Hinsch K. D., Schultz G., Weiler E. W. Identification of GTP-binding proteins in the plasma membrane of higher plants. Biochem Biophys Res Commun. 1988 Oct 31;156(2):954–959. doi: 10.1016/s0006-291x(88)80936-7. [DOI] [PubMed] [Google Scholar]
  4. Breitwieser G. E., Szabo G. Mechanism of muscarinic receptor-induced K+ channel activation as revealed by hydrolysis-resistant GTP analogues. J Gen Physiol. 1988 Apr;91(4):469–493. doi: 10.1085/jgp.91.4.469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown A. M., Birnbaumer L. Ionic channels and their regulation by G protein subunits. Annu Rev Physiol. 1990;52:197–213. doi: 10.1146/annurev.ph.52.030190.001213. [DOI] [PubMed] [Google Scholar]
  6. Brown A. M. Membrane-delimited cell signaling complexes: direct ion channel regulation by G proteins. J Membr Biol. 1993 Jan;131(2):93–104. doi: 10.1007/BF02791318. [DOI] [PubMed] [Google Scholar]
  7. Dolphin A. C., Scott R. H. Calcium channel currents and their inhibition by (-)-baclofen in rat sensory neurones: modulation by guanine nucleotides. J Physiol. 1987 May;386:1–17. doi: 10.1113/jphysiol.1987.sp016518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Fischer R. A. Stomatal opening: role of potassium uptake by guard cells. Science. 1968 May 17;160(3829):784–785. doi: 10.1126/science.160.3829.784. [DOI] [PubMed] [Google Scholar]
  10. Gill D. M., Meren R. ADP-ribosylation of membrane proteins catalyzed by cholera toxin: basis of the activation of adenylate cyclase. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3050–3054. doi: 10.1073/pnas.75.7.3050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gill D. M., Woolkalis M. J. Cholera toxin-catalyzed [32P]ADP-ribosylation of proteins. Methods Enzymol. 1991;195:267–280. doi: 10.1016/0076-6879(91)95172-g. [DOI] [PubMed] [Google Scholar]
  12. Gilman A. G. G proteins: transducers of receptor-generated signals. Annu Rev Biochem. 1987;56:615–649. doi: 10.1146/annurev.bi.56.070187.003151. [DOI] [PubMed] [Google Scholar]
  13. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  14. Hamilton S. L., Codina J., Hawkes M. J., Yatani A., Sawada T., Strickland F. M., Froehner S. C., Spiegel A. M., Toro L., Stefani E. Evidence for direct interaction of Gs alpha with the Ca2+ channel of skeletal muscle. J Biol Chem. 1991 Oct 15;266(29):19528–19535. [PubMed] [Google Scholar]
  15. Hedrich R., Busch H., Raschke K. Ca2+ and nucleotide dependent regulation of voltage dependent anion channels in the plasma membrane of guard cells. EMBO J. 1990 Dec;9(12):3889–3892. doi: 10.1002/j.1460-2075.1990.tb07608.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kopf G. S., Woolkalis M. J. ADP-ribosylation of G proteins with pertussis toxin. Methods Enzymol. 1991;195:257–266. doi: 10.1016/0076-6879(91)95171-f. [DOI] [PubMed] [Google Scholar]
  17. Kuo C. C., Bean B. P. G-protein modulation of ion permeation through N-type calcium channels. Nature. 1993 Sep 16;365(6443):258–262. doi: 10.1038/365258a0. [DOI] [PubMed] [Google Scholar]
  18. Kurachi Y., Nakajima T., Sugimoto T. On the mechanism of activation of muscarinic K+ channels by adenosine in isolated atrial cells: involvement of GTP-binding proteins. Pflugers Arch. 1986 Sep;407(3):264–274. doi: 10.1007/BF00585301. [DOI] [PubMed] [Google Scholar]
  19. Lee H. J., Tucker E. B., Crain R. C., Lee Y. Stomatal Opening Is Induced in Epidermal Peels of Commelina communis L. by GTP Analogs or Pertussis Toxin. Plant Physiol. 1993 May;102(1):95–100. doi: 10.1104/pp.102.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Li W., Assmann S. M. Characterization of a G-protein-regulated outward K+ current in mesophyll cells of vicia faba L. Proc Natl Acad Sci U S A. 1993 Jan 1;90(1):262–266. doi: 10.1073/pnas.90.1.262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Luan S., Li W., Rusnak F., Assmann S. M., Schreiber S. L. Immunosuppressants implicate protein phosphatase regulation of K+ channels in guard cells. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2202–2206. doi: 10.1073/pnas.90.6.2202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ma H., Yanofsky M. F., Meyerowitz E. M. Molecular cloning and characterization of GPA1, a G protein alpha subunit gene from Arabidopsis thaliana. Proc Natl Acad Sci U S A. 1990 May;87(10):3821–3825. doi: 10.1073/pnas.87.10.3821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Moss J., Vaughan M. ADP-ribosylation of guanyl nucleotide-binding regulatory proteins by bacterial toxins. Adv Enzymol Relat Areas Mol Biol. 1988;61:303–379. doi: 10.1002/9780470123072.ch6. [DOI] [PubMed] [Google Scholar]
  24. Schroeder J. I., Raschke K., Neher E. Voltage dependence of K channels in guard-cell protoplasts. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4108–4112. doi: 10.1073/pnas.84.12.4108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Warpeha K. M., Hamm H. E., Rasenick M. M., Kaufman L. S. A blue-light-activated GTP-binding protein in the plasma membranes of etiolated peas. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):8925–8929. doi: 10.1073/pnas.88.20.8925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yatani A., Codina J., Imoto Y., Reeves J. P., Birnbaumer L., Brown A. M. A G protein directly regulates mammalian cardiac calcium channels. Science. 1987 Nov 27;238(4831):1288–1292. doi: 10.1126/science.2446390. [DOI] [PubMed] [Google Scholar]

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