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. 1996 Apr;8(4):701–711. doi: 10.1105/tpc.8.4.701

Cloning and functional expression of a plant voltage-dependent chloride channel.

C Lurin 1, D Geelen 1, H Barbier-Brygoo 1, J Guern 1, C Maurel 1
PMCID: PMC161130  PMID: 8624442

Abstract

Plant cell membrane anion channels participate in basic physiological functions, such as cell volume regulation and signal transduction. However, nothing is known about their molecular structure. Using a polymerase chain reaction strategy, we have cloned a tobacco cDNA (CIC-Nt1) encoding a 780-amino acid protein with several putative transmembrane domains. CIC-Nt1 displays 24 to 32% amino acid identity with members of the animal voltage-dependent chloride channel (CIC) family, whose archetype is CIC-0 from the Torpedo marmorata electric organ. Injection of CIC-Nt1 complementary RNA into Xenopus oocytes elicited slowly activating inward currents upon membrane hyperpolarization more negative than -120 mV. These currents were carried mainly by anions, modulated by extracellular anions, and totally blocked by 10 mM extracellular calcium. The identification of CIC-Nt1 extends the CIC family to higher plants and provides a molecular probe for the study of voltage-dependent anion channels in plants.

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Selected References

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  1. Abrami L., Simon M., Rousselet G., Berthonaud V., Buhler J. M., Ripoche P. Sequence and functional expression of an amphibian water channel, FA-CHIP: a new member of the MIP family. Biochim Biophys Acta. 1994 Jun 1;1192(1):147–151. doi: 10.1016/0005-2736(94)90155-4. [DOI] [PubMed] [Google Scholar]
  2. Adachi S., Uchida S., Ito H., Hata M., Hiroe M., Marumo F., Sasaki S. Two isoforms of a chloride channel predominantly expressed in thick ascending limb of Henle's loop and collecting ducts of rat kidney. J Biol Chem. 1994 Jul 1;269(26):17677–17683. [PubMed] [Google Scholar]
  3. Cavener D. R., Ray S. C. Eukaryotic start and stop translation sites. Nucleic Acids Res. 1991 Jun 25;19(12):3185–3192. doi: 10.1093/nar/19.12.3185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Felle H. H. The H+/Cl- Symporter in Root-Hair Cells of Sinapis alba (An Electrophysiological Study Using Ion-Selective Microelectrodes). Plant Physiol. 1994 Nov;106(3):1131–1136. doi: 10.1104/pp.106.3.1131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fujita N., Mori H., Yura T., Ishihama A. Systematic sequencing of the Escherichia coli genome: analysis of the 2.4-4.1 min (110,917-193,643 bp) region. Nucleic Acids Res. 1994 May 11;22(9):1637–1639. doi: 10.1093/nar/22.9.1637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Greene J. R., Brown N. H., DiDomenico B. J., Kaplan J., Eide D. J. The GEF1 gene of Saccharomyces cerevisiae encodes an integral membrane protein; mutations in which have effects on respiration and iron-limited growth. Mol Gen Genet. 1993 Dec;241(5-6):542–553. doi: 10.1007/BF00279896. [DOI] [PubMed] [Google Scholar]
  8. Gründer S., Thiemann A., Pusch M., Jentsch T. J. Regions involved in the opening of CIC-2 chloride channel by voltage and cell volume. Nature. 1992 Dec 24;360(6406):759–762. doi: 10.1038/360759a0. [DOI] [PubMed] [Google Scholar]
  9. Hedrich R., Becker D. Green circuits--the potential of plant specific ion channels. Plant Mol Biol. 1994 Dec;26(5):1637–1650. doi: 10.1007/BF00016494. [DOI] [PubMed] [Google Scholar]
  10. Hedrich R., Marten I. Malate-induced feedback regulation of plasma membrane anion channels could provide a CO2 sensor to guard cells. EMBO J. 1993 Mar;12(3):897–901. doi: 10.1002/j.1460-2075.1993.tb05730.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jentsch T. J., Günther W., Pusch M., Schwappach B. Properties of voltage-gated chloride channels of the ClC gene family. J Physiol. 1995 Jan;482:19S–25S. doi: 10.1113/jphysiol.1995.sp020560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jentsch T. J., Steinmeyer K., Schwarz G. Primary structure of Torpedo marmorata chloride channel isolated by expression cloning in Xenopus oocytes. Nature. 1990 Dec 6;348(6301):510–514. doi: 10.1038/348510a0. [DOI] [PubMed] [Google Scholar]
  13. Kawasaki M., Suzuki M., Uchida S., Sasaki S., Marumo F. Stable and functional expression of the CIC-3 chloride channel in somatic cell lines. Neuron. 1995 Jun;14(6):1285–1291. doi: 10.1016/0896-6273(95)90275-9. [DOI] [PubMed] [Google Scholar]
  14. Kawasaki M., Uchida S., Monkawa T., Miyawaki A., Mikoshiba K., Marumo F., Sasaki S. Cloning and expression of a protein kinase C-regulated chloride channel abundantly expressed in rat brain neuronal cells. Neuron. 1994 Mar;12(3):597–604. doi: 10.1016/0896-6273(94)90215-1. [DOI] [PubMed] [Google Scholar]
  15. Kowdley G. C., Ackerman S. J., John J. E., 3rd, Jones L. R., Moorman J. R. Hyperpolarization-activated chloride currents in Xenopus oocytes. J Gen Physiol. 1994 Feb;103(2):217–230. doi: 10.1085/jgp.103.2.217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  17. Marchuk D., Drumm M., Saulino A., Collins F. S. Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products. Nucleic Acids Res. 1991 Mar 11;19(5):1154–1154. doi: 10.1093/nar/19.5.1154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Maurel C., Kado R. T., Guern J., Chrispeels M. J. Phosphorylation regulates the water channel activity of the seed-specific aquaporin alpha-TIP. EMBO J. 1995 Jul 3;14(13):3028–3035. doi: 10.1002/j.1460-2075.1995.tb07305.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Maurel C., Reizer J., Schroeder J. I., Chrispeels M. J. The vacuolar membrane protein gamma-TIP creates water specific channels in Xenopus oocytes. EMBO J. 1993 Jun;12(6):2241–2247. doi: 10.1002/j.1460-2075.1993.tb05877.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Middleton R. E., Pheasant D. J., Miller C. Purification, reconstitution, and subunit composition of a voltage-gated chloride channel from Torpedo electroplax. Biochemistry. 1994 Nov 15;33(45):13189–13198. doi: 10.1021/bi00249a005. [DOI] [PubMed] [Google Scholar]
  21. Murray E. E., Lotzer J., Eberle M. Codon usage in plant genes. Nucleic Acids Res. 1989 Jan 25;17(2):477–498. doi: 10.1093/nar/17.2.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pantoja O., Gelli A., Blumwald E. Characterization of Vacuolar Malate and K Channels under Physiological Conditions. Plant Physiol. 1992 Nov;100(3):1137–1141. doi: 10.1104/pp.100.3.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Plant P. J., Gelli A., Blumwald E. Vacuolar chloride regulation of an anion-selective tonoplast channel. J Membr Biol. 1994 May;140(1):1–12. doi: 10.1007/BF00234481. [DOI] [PubMed] [Google Scholar]
  24. Rosenthal E. R., Guidotti G. Reconstitution, identification, and purification of the Torpedo californica electroplax chloride channel complex. Biochim Biophys Acta. 1994 May 11;1191(2):256–266. doi: 10.1016/0005-2736(94)90176-7. [DOI] [PubMed] [Google Scholar]
  25. Schroeder J. I. Anion channels as central mechanisms for signal transduction in guard cells and putative functions in roots for plant-soil interactions. Plant Mol Biol. 1995 Jun;28(3):353–361. doi: 10.1007/BF00020385. [DOI] [PubMed] [Google Scholar]
  26. Shimbo K., Brassard D. L., Lamb R. A., Pinto L. H. Viral and cellular small integral membrane proteins can modify ion channels endogenous to Xenopus oocytes. Biophys J. 1995 Nov;69(5):1819–1829. doi: 10.1016/S0006-3495(95)80052-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Steinmeyer K., Ortland C., Jentsch T. J. Primary structure and functional expression of a developmentally regulated skeletal muscle chloride channel. Nature. 1991 Nov 28;354(6351):301–304. doi: 10.1038/354301a0. [DOI] [PubMed] [Google Scholar]
  28. Thiemann A., Gründer S., Pusch M., Jentsch T. J. A chloride channel widely expressed in epithelial and non-epithelial cells. Nature. 1992 Mar 5;356(6364):57–60. doi: 10.1038/356057a0. [DOI] [PubMed] [Google Scholar]
  29. Thomine S., Zimmermann S., Guern J., Barbier-Brygoo H. ATP-Dependent Regulation of an Anion Channel at the Plasma Membrane of Protoplasts from Epidermal Cells of Arabidopsis Hypocotyls. Plant Cell. 1995 Dec;7(12):2091–2100. doi: 10.1105/tpc.7.12.2091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Uchida S., Sasaki S., Nitta K., Uchida K., Horita S., Nihei H., Marumo F. Localization and functional characterization of rat kidney-specific chloride channel, ClC-K1. J Clin Invest. 1995 Jan;95(1):104–113. doi: 10.1172/JCI117626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. van Slegtenhorst M. A., Bassi M. T., Borsani G., Wapenaar M. C., Ferrero G. B., de Conciliis L., Rugarli E. I., Grillo A., Franco B., Zoghbi H. Y. A gene from the Xp22.3 region shares homology with voltage-gated chloride channels. Hum Mol Genet. 1994 Apr;3(4):547–552. doi: 10.1093/hmg/3.4.547. [DOI] [PubMed] [Google Scholar]

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