Arabidopsis AtCNGC10 rescues potassium channel mutants of E. coli, yeast and Arabidopsis and is regulated by calcium / calmodulin and cyclic GMP in E. coli
Xinli Li A , Tamás Borsics A , H. Michael Harrington A and David A. Christopher A BA University of Hawaii, Department of Molecular Biosciences and Bioengineering 1955 East–West Road, Agsciences 218 Honolulu, HI 96822, Hawaii.
B Corresponding author. Email: dchr@hawaii.edu
Functional Plant Biology 32(7) 643-653 https://doi.org/10.1071/FP04233
Submitted: 14 December 2004 Accepted: 15 April 2005 Published: 7 July 2005
Abstract
We have isolated and characterised AtCNGC10, one of the 20 members of the family of cyclic nucleotide (CN)-gated and calmodulin (CaM)-regulated channels (CNGCs) from Arabidopsis thaliana (L.) Heynh. AtCNGC10 bound CaM in a C-terminal subregion that contains a basic amphiphillic structure characteristic of CaM-binding proteins and that also overlaps with the predicted CN-binding domain. AtCNGC10 is insensitive to the broad-range K+ channel blocker, tetraethylammonium, and lacks a typical K+-signature motif. However, AtCNGC10 complemented K+ channel uptake mutants of Escherichia coli (LB650), yeast (Saccharomyces cerevisiae CY162) and Arabidopsis (akt1-1). Sense 35S-AtCNGC10 transformed into the Arabidopsis akt1-1 mutant, grew 1.7-fold better on K+-limited medium relative to the vector control. Coexpression of CaM and AtCNGC10 in E. coli showed that Ca2+ / CaM inhibited cell growth by 40%, while cGMP reversed the inhibition by Ca2+ / CaM, in a AtCNGC10-dependent manner. AtCNGC10 did not confer tolerance to Cs+ in E. coli, however, it confers tolerance to toxic levels of Na+ and Cs+ in the yeast K+ uptake mutant grown on low K+ medium. Antisense AtCNGC10 plants had 50% less potassium than wild type Columbia. Taken together, the studies from three evolutionarily diverse species demonstrated a role for the CaM-binding channel, AtCNGC10, in mediating the uptake of K+ in plants.
Keywords: Arabidopsis, calmodulin, CNGC, complementation, cyclic nucleotide, K+ channel.
Acknowledgments
This research was supported by funds from the USA Department of Energy grant No. DE-FG02–03ER15395 to DAC.
Anderson JA,
Huprikar SS,
Kochian LV,
Lucas WJ, Gaber RF
(1992) Functional expression of a probable Arabidopsis thaliana potassium channel in Saccharomyces cerevisiae.
Proceedings of the National Academy of Sciences USA 89, 3736–3740.
Arazi T,
Sunkar R,
Kaplan B, Fromm H
(1999) A tobacco plasma membrane calmodulin-binding transporter confers Ni2+ tolerance and Pb2+ hypersensitivity in transgenic plants. The Plant Journal 20, 171–182.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Balagué C,
Lin B,
Alcon C,
Flottes G,
Malmström S,
Köhler C,
Neuhaus G,
Pelletier G,
Gaymard F, Roby D
(2003) HLM1, an essential signaling component in the hypersensitive response, is a member of the cyclic nucleotide-gated channel ion channel family. The Plant Cell 15, 365–379.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Botsford JL, Harman JG
(1992) Cyclic AMP in prokaryotes. Microbiology and Molecular Biology Reviews 56, 100–122.
Burgess WH,
Watterson DM, Van Eldik LJ
(1984) Identification of calmodulin binding proteins in chicken embryo fibroblasts. Journal of Cell Biology 46, 466–469.
Bush DS
(1995) Calcium regulation in plant cells and its role in signaling. Annual Review of Plant Physiology and Plant Molecular Biology 46, 95–122.
| Crossref | GoogleScholarGoogle Scholar |
Cao Y,
Ward JM,
Kelly WB,
Ichida AM,
Gaber RF,
Anderson JA,
Uozumi N,
Schroeder JI, Crawford NM
(1995) Multiple genes, tissue specificity, and expression-dependent modulation contribute to the functional diversity of potassium channels in Arabidopsis thaliana.
Plant Physiology 109, 1093–1106.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Chan CWM,
Schorrak LM,
Smith RK,
Bent AF, Sussman MR
(2003) A cyclic nucleotide-gated ion channel, CNGC2, is crucial for plant development and adaptation to calcium stress. Plant Physiology 132, 728–731.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Clough SJ,
Fengler KA,
Lippok B,
Smith RK,
Yu I-C, Bent AF
(2000) The Arabidopsis dnd1 ‘defense, no death’ gene encodes a mutated cyclic nucleotide-gated ion channel. Proceedings of the National Academy of Sciences USA 97, 9323–9328.
| Crossref | GoogleScholarGoogle Scholar |
Doyle DA,
Morais-Cabral J,
Pfuetzner RA,
Kuo A,
Gulbis JM,
Cohen SL,
Chait BT, MacKinnon R
(1998) The structure of the potassium channel: molecular basis of K+ conduction and selectivity. Science 280, 69–77.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Eismann E,
Muller F,
Heinemann SH, Kaupp UB
(1994) A single negative charge within the pore region of a cGMP gated channel controls rectification, Ca2+ blockage, and ionic selectivity. Proceedings of the National Academy of Sciences USA 91, 1109–1113.
Fairbairn DJ,
Liu W,
Schachtman DP,
Gomez-Gallego S,
Day SR, Teasdale RD
(2000) Characterisation of two distinct HKT1-like potassium transporters from Eucalyptus camaldulensis.
Plant Molecular Biology Erratum in: Plant Molecular Biology (2001) 45, 245. 43, 515–525.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Finn JT,
Krautwurst D,
Schroeder JE,
Chen T-Y,
Reed RR, Yau K-W
(1998) Functional co-assembly among subunits of cyclic-nucleotide-activated, nonselective cation channels, and across species from nematode to human. Biophysical Journal 74, 1333–1345.
| PubMed |
Fox TC, Guerinot ML
(1998) Molecular biology of cation transport in plants. Annual Review of Plant Physiology and Plant Molecular Biology 49, 669–696.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Gaymard F,
Cerutti M,
Horeau C,
Lemaillet G,
Urbach S,
Ravallec M,
Devauchelle G,
Sentenac H, Thibaud JB
(1996) The baculovirus / insect cell system as an alternative to Xenopus oocytes. First characterization of the AKT1 K+ channel from Arabidopsis thaliana.
Journal of Biological Chemistry 271, 22863–22870.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Gaymard F,
Pilot G,
Lacombe B,
Bouchez D,
Bruneau D,
Boucherez J,
Michaux-Ferriere N,
Thibaud JB, Sentenac H
(1998) Identification and disruption of a plant shaker-like outward channel involved in K+ release into the xylem sap. Cell 94, 647–655.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Gralla, JD ,
and
Collado-Vides, J (1996). Organization and function of transcription regulatory elements. In ‘ and ’. pp. 1232–1245. (American Society Microbiology Press: Washington DC)
Grunwald ME,
Zhong H, Yau K-W
(1999) Molecular determinants of the modulation of cyclic nucleotide-activated channels by calmodulin. Proceedings of the National Academy of Sciences USA 96, 13444–13449.
| Crossref | GoogleScholarGoogle Scholar |
Hennegan KP, Danna KJ
(1998) pBIN20: An improved vector for shape Agrobacterium-mediated transformation. Plant Molecular Biology Reporter 16, 129–131.
| Crossref | GoogleScholarGoogle Scholar |
Herdman M, Elmorjani K
(1988) Cyclic nucleotides. Methods in Enzymology 167, 584–591.
Hirsch RE,
Lewis BD,
Spalding EP, Sussman MR
(1998) A role for the AKT1 potassium channel in plant nutrition. Science 280, 918–920.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Hoshi T
(1995) Regulation of voltage-dependence of the KAT1 channel by intracellular factors. Journal of General Physiology 105, 309–328.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Hua BG,
Mercier RW,
Zielinski RE, Berkowitz GA
(2003) Functional interaction of calmodulin with a plant cyclic nucleotide gated cation channel. Plant Physiology and Biochemistry 41, 945–954.
| Crossref | GoogleScholarGoogle Scholar |
Ketchum KA, Slayman CW
(1996) Isolation of an ion channel gene from Arabidopsis thaliana using the H5 signature sequence from voltage-dependent K+ channels. FEBS Letters 378, 19–26.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Ko CH, Gaber RF
(1991) TRK1 and TRK2 encode structurally related K+ transporters in Saccharomyces cerevisiae.
Molecular and Cellular Biology 11, 4266–4273.
| PubMed |
Köhler C, Neuhaus G
(2000) Characterization of calmodulin binding to cyclic nucleotide-gated ion channels from Arabidopsis thaliana.
FEBS Letters 471, 133–136.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Köhler C,
Merkle T, Neuhaus G
(1999) Characterization of a novel gene family of putative cyclic nucleotide- and calmodulin-regulated ion channels in Arabidopsis thaliana.
The Plant Journal 18, 97–104.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Kolb A,
Busby S,
Buc H,
Garges S, Adhya S
(1993) Transcriptional regulation by cAMP and its receptor protein. Annual Review of Biochemistry 62, 749–795.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Laemmli UK
(1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–
.
| PubMed |
Leng Q,
Mercier RW,
Yao W, Berkowitz GA
(1999) Cloning and first functional characterization of a plant cyclic nucleotide-gated cation channel. Plant Physiology 121, 753–761.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Leng Q,
Mercier RW,
Hua B-G,
Fromm H, Berkowitz GA
(2002) Electrophysiological analysis of cloned cyclic nucleotide-gated ion channels. Plant Physiology 128, 400–410.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Li W,
Luan S,
Schreiber SL, Assmann SM
(1994) Cyclic AMP stimulates K+ channel activity in mesophyll cells of Vicia faba L. Plant Physiology 106, 957–961.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Liu M,
Chen TY,
Ahamed B,
Li J, Yau K-W
(1994) Calcium-calmodulin modification of the olfactory cyclic nucleotide-gated cation channel. Science 266, 1348–1354.
| PubMed |
Lu YT, Harrington HM
(1994) Isolation of tobacco cDNA clones encoding-binding proteins and characterization of calmodulin-binding domain. Plant Physiology and Biochemistry 32, 413–422.
Maathuis FJ,
Ichida AM,
Sanders D, Schroeder JI
(1997) Roles of higher plant K+ channels. Plant Physiology 114, 1141–1149.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Mäser P,
Thomine S,
Schroeder JI,
Ward JM, Hirschi K , et al.
(2001) Phylogenetic relationships with cation transporter families of Arabidopsis.
Plant Physiology 126, 1646–1667.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Nakamura RL,
Anderson JA, Gaber RF
(1997) Determination of key structural requirements of a K+ channel pore. Journal of Biological Chemistry 272, 1011–1018.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Navarre C, Goffeau A
(2000) Membrane hyperpolarization and salt sensitivity induced by deletion of PMP3, a highly conserved small protein of yeast plasma membrane. EMBO Journal 19, 2515–2524.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
O’Neil KT, DeGrado WF
(1990) How calmodulin binds it targets: sequence independent recognition of amphiphilic alpha-helices. Trends in Biochemical Sciences 15, 59–64.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Pilot G,
Gaymard F,
Mouline K,
Cherel I, Sentenac H
(2003) Regulated expression of Arabidopsis shaker K+ channel genes involved in K+ uptake and distribution in the plant. Plant Molecular Biology 51, 773–787.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Reintanz B,
Szyroki A,
Ivashikina N,
Ache P,
Godde M,
Becker D,
Palme K, Hedrich R
(2002) AtKC1, a silent Arabidopsis potassium channel α-subunit modulates root hair K+-influx. Proceedings of the National Academy of Sciences USA 99, 4079–4084.
| Crossref | GoogleScholarGoogle Scholar |
Root MJ, MacKinnon R
(1993) Identification of an external divalent cation-binding site in the pore of a cGMP-activated channel. Neuron 11, 459–466.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Schachtman DP
(2000) Molecular insights into the structure and function of plant K+ transport mechanisms. Biochimica et Biophysica Acta 1465, 127–139.
| PubMed |
Schlösser A,
Meldorf M,
Stumpe S,
Bakker EP, Epstein W
(1995) TrkH and its homolog, TrkG, determine the specificity and kinetics of cation transport by the Trk system of Escherichia coli.
Journal of Bacteriology 177, 1908–1910.
| PubMed |
Schuurink RC,
Shartzer SF,
Fath A, Jones RL
(1998) Characterization of a calmodulin-binding transporter from plasma membrane of barley aleurone. Proceedings of the National Academy USA 95, 1944–1949.
| Crossref | GoogleScholarGoogle Scholar |
Sentenac H,
Bonneaud N,
Minet M,
Lacroute F,
Salmon JM,
Gaymard F, Grignon C
(1992) Cloning and expression in yeast of a plant potassium ion transport system. Science 256, 663–665.
| PubMed |
Stumpe S, Bakker EP
(1997) Requirement of a large K+-uptake capacity and of extracytoplasmic protease activity for protamine resistance of Escherichia coli.
Archives of Microbiology 167, 126–136.
| Crossref | GoogleScholarGoogle Scholar |
Talke IN,
Blaudez D,
Maathuis FJ, Sanders D
(2003) CNGCs: prime targets of plant cyclic nucleotide signalling? Trends in Plant Science 8, 286–293.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Tang HX,
Vasconcelos AC, Berkowitz GA
(1996) Physical association of KAB1 with plant K+ channel alpha subunits. The Plant Cell 8, 1545–1553.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Trudeau MC, Zagotta WN
(2002) Mechanism of calcium / calmodulin inhibition of rod cyclic nucleotide-gated channels. Proceedings of the National Academy of Sciences USA 99, 8424–8429.
| Crossref | GoogleScholarGoogle Scholar |
Varnum MD, Zagotta WN
(1997) Interdomain interactions underlying activation of cyclic nucleotide-gated channels. Science 278, 110–113.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Vogler AP, Lengeler JW
(1987) Indirect role of adenylate cyclase and cyclic AMP in chemotaxis to phosphotransferase system carbohydrates in Escherichia coli K-12. Journal of Bacteriology 169, 593–599.
| PubMed |
Zielinski RE
(1998) Calmodulin and calmodulin-binding proteins in plants. Annual Review of Plant Physiology and Plant Molecular Biology 49, 697–725.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
