title: Identification of cyclic GMP-activated nonselective Ca2+-permeable cation channels and associated CNGC5 and CNGC6 genes in Arabidopsis guard cells.
abstract: Cytosolic Ca(2+) in guard cells plays an important role in stomatal movement responses to environmental stimuli. These cytosolic Ca(2+) increases result from Ca(2+) influx through Ca(2+)-permeable channels in the plasma membrane and Ca(2+) release from intracellular organelles in guard cells. However, the genes encoding defined plasma membrane Ca(2+)-permeable channel activity remain unknown in guard cells and, with some exceptions, largely unknown in higher plant cells. Here, we report the identification of two Arabidopsis (Arabidopsis thaliana) cation channel genes, CNGC5 and CNGC6, that are highly expressed in guard cells. Cytosolic application of cyclic GMP (cGMP) and extracellularly applied membrane-permeable 8-Bromoguanosine 3',5'-cyclic monophosphate-cGMP both activated hyperpolarization-induced inward-conducting currents in wild-type guard cells using Mg(2+) as the main charge carrier. The cGMP-activated currents were strongly blocked by lanthanum and gadolinium and also conducted Ba(2+), Ca(2+), and Na(+) ions. cngc5 cngc6 double mutant guard cells exhibited dramatically impaired cGMP-activated currents. In contrast, mutations in CNGC1, CNGC2, and CNGC20 did not disrupt these cGMP-activated currents. The yellow fluorescent protein-CNGC5 and yellow fluorescent protein-CNGC6 proteins localize in the cell periphery. Cyclic AMP activated modest inward currents in both wild-type and cngc5cngc6 mutant guard cells. Moreover, cngc5 cngc6 double mutant guard cells exhibited functional abscisic acid (ABA)-activated hyperpolarization-dependent Ca(2+)-permeable cation channel currents, intact ABA-induced stomatal closing responses, and whole-plant stomatal conductance responses to darkness and changes in CO2 concentration. Furthermore, cGMP-activated currents remained intact in the growth controlled by abscisic acid2 and abscisic acid insensitive1 mutants. This research demonstrates that the CNGC5 and CNGC6 genes encode unique cGMP-activated nonselective Ca(2+)-permeable cation channels in the plasma membrane of Arabidopsis guard cells.
authors.affiliation: National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
authors.forename: Yong-Fei
authors.initials: YF
authors.lastname: Wang
authors.forename: Shintaro
authors.initials: S
authors.lastname: Munemasa
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Nishimura
authors.forename: Hui-Min
authors.initials: HM
authors.lastname: Ren
authors.forename: Nadia
authors.initials: N
authors.lastname: Robert
authors.forename: Michelle
authors.initials: M
authors.lastname: Han
authors.forename: Irina
authors.initials: I
authors.lastname: Puzõrjova
authors.forename: Hannes
authors.initials: H
authors.lastname: Kollist
authors.forename: Stephen
authors.initials: S
authors.lastname: Lee
authors.forename: Izumi
authors.initials: I
authors.lastname: Mori
authors.forename: Julian I
authors.initials: JI
authors.lastname: Schroeder
mesh_terms: D000040:Abscisic Acid / Q000494:pharmacology; D017360:Arabidopsis / Q000166:cytology / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000378:metabolism*; D002245:Carbon Dioxide / Q000494:pharmacology; D002412:Cations; D002463:Cell Membrane Permeability / Q000187:drug effects*; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000494:pharmacology*; D054815:Cyclic Nucleotide-Gated Cation Channels / Q000235:genetics* / Q000378:metabolism; D060146:Ecotype; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017343:Genes, Plant / Q000235:genetics; D015640:Ion Channel Gating / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D008027:Light; D009154:Mutation / Q000235:genetics; D054046:Plant Stomata / Q000166:cytology* / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D011523:Protoplasts / Q000187:drug effects / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D013997:Time Factors; D014026:Nicotiana / Q000187:drug effects / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; C588161:CNGC5 protein, Arabidopsis; C588162:CNGC6 protein, Arabidopsis; D002121:Calcium Channel Blockers; D015220:Calcium Channels; D002412:Cations; D054815:Cyclic Nucleotide-Gated Cation Channels; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D002245:Carbon Dioxide; C016276:8-bromocyclic GMP; D000040:Abscisic Acid; D006152:Cyclic GMP; D002118:Calcium
references.citation: Plant J. 2000 Nov;24(4):533-42
references.pmid: 11115134
references.citation: Plant Cell. 1994 May;6(5):685-694
references.pmid: 12244254
references.citation: Plant Cell Physiol. 2013 Apr;54(4):573-84
references.pmid: 23385145
references.citation: Plant Cell. 2006 Apr;18(4):935-54
references.pmid: 16531497
references.citation: Plant Physiol. 2013 Jul;162(3):1681-93
references.pmid: 23735507
references.citation: Plant J. 2002 Oct;32(2):185-94
references.pmid: 12383084
references.citation: Plant Physiol. 2007 Jan;143(1):28-37
references.pmid: 17142476
references.citation: Plant Cell Physiol. 2012 Dec;53(12):2047-59
references.pmid: 23054389
references.citation: Plant Cell. 2007 Oct;19(10):3019-36
references.pmid: 17921317
references.citation: Plant Cell. 2001 Nov;13(11):2513-23
references.pmid: 11701885
references.citation: Plant Mol Biol. 2007 Dec;65(6):747-61
references.pmid: 17885810
references.citation: Plant Cell. 2012 Aug;24(8):3167
references.pmid: 22904149
references.citation: Plant Cell. 2012 Jun;24(6):2546-61
references.pmid: 22730405
references.citation: Proc Natl Acad Sci U S A. 2000 Aug 1;97(16):9323-8
references.pmid: 10900264
references.citation: New Phytol. 2011 Jul;191(1):57-69
references.pmid: 21371039
references.citation: Plant Cell. 1994 May;6(5):669-683
references.pmid: 12244253
references.citation: Science. 1998 Oct 9;282(5387):287-90
references.pmid: 9765153
references.citation: Plant J. 2010 Mar;61(5):816-25
references.pmid: 20015065
references.citation: Proc Natl Acad Sci U S A. 1990 Dec;87(23):9305-9
references.pmid: 2174559
references.citation: PLoS Biol. 2006 Oct;4(10):e327
references.pmid: 17032064
references.citation: Curr Biol. 2013 Jan 7;23(1):53-7
references.pmid: 23219726
references.citation: Nature. 2000 Aug 17;406(6797):731-4
references.pmid: 10963598
references.citation: Science. 2001 May 25;292(5521):1486-7
references.pmid: 11379626
references.citation: Plant Physiol. 2007 Mar;143(3):1398-407
references.pmid: 17220365
references.citation: Plant Cell. 2002 Jul;14(7):1649-62
references.pmid: 12119381
references.citation: Proc Natl Acad Sci U S A. 2011 May 10;108(19):E96; author reply E97-8
references.pmid: 21527716
references.citation: J Biol Chem. 2003 Feb 21;278(8):6490-4
references.pmid: 12482758
references.citation: Plant Physiol. 2002 Jun;129(2):469-85
references.pmid: 12068094
references.citation: J Biol Chem. 2007 Jan 19;282(3):1916-24
references.pmid: 17098742
references.citation: FEBS Lett. 2007 May 25;581(12):2237-46
references.pmid: 17321525
references.citation: Plant Physiol. 2013 Feb;161(2):1010-20
references.pmid: 23370720
references.citation: Plant Signal Behav. 2011 Mar;6(3):413-5
references.pmid: 21358265
references.citation: Plant J. 2010 Apr;62(2):265-76
references.pmid: 20088896
references.citation: Plant J. 1999 Apr;18(1):97-104
references.pmid: 10341447
references.citation: Plant Physiol. 1988 Jul;87(3):583-7
references.pmid: 16666189
references.citation: Mol Pharmacol. 2004 Mar;65(3):503-11
references.pmid: 14978228
references.citation: Plant Physiol. 2011 Feb;155(2):963-73
references.pmid: 21177472
references.citation: Plant Cell. 1999 Sep;11(9):1785-98
references.pmid: 10488243
references.citation: Plant Cell. 2004 Mar;16(3):596-615
references.pmid: 14973164
references.citation: Proc Natl Acad Sci U S A. 2010 Apr 27;107(17):8023-8
references.pmid: 20385816
references.citation: Proc Natl Acad Sci U S A. 2000 Apr 25;97(9):4967-72
references.pmid: 10781106
references.citation: Mol Syst Biol. 2010 Jun 8;6:372
references.pmid: 20531402
references.citation: Plant Cell. 1999 Dec;11(12):2283-90
references.pmid: 10590158
references.citation: J Biol Chem. 2004 Aug 20;279(34):35306-12
references.pmid: 15199067
references.citation: Plant J. 2001 May;26(3):351-62
references.pmid: 11439123
references.citation: Plant Cell. 1992 Sep;4(9):1113-1122
references.pmid: 12297670
references.citation: Proc Natl Acad Sci U S A. 2012 Jun 26;109(26):10593-8
references.pmid: 22689970
references.citation: Plant J. 2009 Jul;59(2):207-20
references.pmid: 19302418
references.citation: Plant J. 2012 Jun;70(6):1056-69
references.pmid: 22372427
references.citation: Proc Natl Acad Sci U S A. 2009 Dec 15;106(50):21425-30
references.pmid: 19955405
references.citation: Plant Cell. 2003 Feb;15(2):365-79
references.pmid: 12566578
references.citation: EMBO J. 2011 Apr 20;30(8):1645-58
references.pmid: 21423149
references.citation: Plant Cell. 2012 Aug;24(8):3333-48
references.pmid: 22904147
references.citation: Plant Physiol. 2010 Nov;154(3):1232-43
references.pmid: 20805328
references.citation: Plant Cell. 2012 Apr;24(4):1522-33
references.pmid: 22523205
references.citation: Plant Physiol. 2004 Dec;136(4):3892-904
references.pmid: 15542492
references.citation: Annu Rev Plant Biol. 2010;61:593-620
references.pmid: 20192754
references.citation: Plant Cell. 2006 Oct;18(10):2749-66
references.pmid: 16998070
references.citation: EMBO J. 1994 Jul 1;13(13):2970-5
references.pmid: 8039493
references.citation: J Exp Bot. 2006;57(1):125-38
references.pmid: 16317039
references.citation: Nature. 2008 Mar 27;452(7186):487-91
references.pmid: 18305484
references.citation: Plant Cell Physiol. 2002 Dec;43(12):1473-83
references.pmid: 12514244
references.citation: Annu Rev Physiol. 2009;71:59-82
references.pmid: 18842100
references.citation: BMC Plant Biol. 2009 Nov 27;9:140
references.pmid: 19943938
references.citation: Ann Bot. 2012 Jan;109(1):5-17
references.pmid: 21994053
references.citation: Proc Natl Acad Sci U S A. 2009 Dec 15;106(50):21419-24
references.pmid: 19955427
references.citation: Proc Natl Acad Sci U S A. 1998 Jun 23;95(13):7819-24
references.pmid: 9636234
references.citation: Proc Natl Acad Sci U S A. 2000 Oct 24;97(22):12361-8
references.pmid: 11027317
references.citation: Neuron. 1995 Jan;14(1):177-83
references.pmid: 7530019
references.citation: Proc Natl Acad Sci U S A. 2006 May 9;103(19):7506-11
references.pmid: 16651523
references.citation: Proc Natl Acad Sci U S A. 1998 Apr 14;95(8):4778-83
references.pmid: 9539815
references.citation: Plant Cell. 2002 Dec;14(12):3089-99
references.pmid: 12468729
references.citation: Plant Methods. 2008 Feb 19;4:6
references.pmid: 18284694
references.citation: Proc Natl Acad Sci U S A. 2000 Aug 15;97(17):9801-6
references.pmid: 10920194
references.citation: Plant Physiol. 2012 May;159(1):40-6
references.pmid: 22447719
references.citation: Curr Opin Plant Biol. 2004 Oct;7(5):537-46
references.pmid: 15337096
references.citation: Plant Physiol. 2002 Feb;128(2):400-10
references.pmid: 11842144
references.citation: Proc Natl Acad Sci U S A. 2010 Dec 7;107(49):21193-8
references.pmid: 21088220
references.citation: Plant Physiol. 2010 Oct;154(2):463-6
references.pmid: 20921164
references.citation: Proc Natl Acad Sci U S A. 2007 Sep 4;104(36):14531-6
references.pmid: 17726111
references.citation: Science. 2011 Apr 22;332(6028):434-7
references.pmid: 21415319
references.citation: EMBO J. 2003 Jun 2;22(11):2623-33
references.pmid: 12773379
references.citation: J Microsc. 2004 May;214(Pt 2):159-73
references.pmid: 15102063
references.citation: Proc Natl Acad Sci U S A. 1998 Feb 17;95(4):1944-9
references.pmid: 9465122
references.citation: PLoS One. 2013;8(2):e55277
references.pmid: 23424627
references.citation: Plant Cell. 2007 Mar;19(3):1081-95
references.pmid: 17384171
references.citation: Plant J. 1999 Oct;20(2):171-82
references.pmid: 10571877
references.citation: Physiol Plant. 2010 Jul 1;139(3):303-12
references.pmid: 20210874
references.citation: Annu Rev Plant Biol. 2010;61:561-91
references.pmid: 20192751
references.citation: Nature. 2002 Feb 28;415(6875):977-83
references.pmid: 11875555
references.citation: Physiol Plant. 2008 Nov;134(3):499-507
references.pmid: 18823330
references.citation: Plant Physiol. 1999 Jan;119(1):277-88
references.pmid: 9880370