A family of putative chloride channels from Arabidopsis and functional complementation of a yeast strain with a CLC gene disruption

TY - JOUR

T1 - A family of putative chloride channels from Arabidopsis and functional complementation of a yeast strain with a CLC gene disruption

AU - Hechenberger, M

AU - Schwappach, B

AU - Fischer, W N

AU - Frommer, W B

AU - Jentsch, T J

AU - Steinmeyer, K

PY - 1996/12/27

Y1 - 1996/12/27

N2 - We have cloned four novel members of the CLC family of chloride channels from Arabidopsis thaliana. The four plant genes are homologous to a recently isolated chloride channel gene from tobacco (CLC-Nt1; Lurin, C., Geelen, D., Barbier-Brygoo, H., Guern, J., and Maurel, C. (1996) Plant Cell 8, 701-711) and are about 30% identical in sequence to the most closely related CLC-6 and CLC-7 putative chloride channels from mammalia. AtCLC transcripts are broadly expressed in the plant. Similarly, antibodies against the AtCLC-d protein detected the protein in all tissues, but predominantly in the silique. AtCLC-a and AtCLC-b are highly homologous to each other ( approximately 87% identity), while being approximately 50% identical to either AtCLC-c or AtCLC-d. None of the four cDNAs elicited chloride currents when expressed in Xenopus oocytes, either singly or in combination. Among these genes, only AtCLC-d could functionally substitute for the single yeast CLC protein, restoring iron-limited growth of a strain disrupted for this gene. Introduction of disease causing mutations, identified in human CLC genes, abolished this capacity. Consistent with a similar function of both proteins, the green fluorescent protein-tagged AtCLC-d protein showed the identical localization pattern as the yeast ScCLC protein. This suggests that in Arabidopsis AtCLC-d functions as an intracellular chloride channel.

AB - We have cloned four novel members of the CLC family of chloride channels from Arabidopsis thaliana. The four plant genes are homologous to a recently isolated chloride channel gene from tobacco (CLC-Nt1; Lurin, C., Geelen, D., Barbier-Brygoo, H., Guern, J., and Maurel, C. (1996) Plant Cell 8, 701-711) and are about 30% identical in sequence to the most closely related CLC-6 and CLC-7 putative chloride channels from mammalia. AtCLC transcripts are broadly expressed in the plant. Similarly, antibodies against the AtCLC-d protein detected the protein in all tissues, but predominantly in the silique. AtCLC-a and AtCLC-b are highly homologous to each other ( approximately 87% identity), while being approximately 50% identical to either AtCLC-c or AtCLC-d. None of the four cDNAs elicited chloride currents when expressed in Xenopus oocytes, either singly or in combination. Among these genes, only AtCLC-d could functionally substitute for the single yeast CLC protein, restoring iron-limited growth of a strain disrupted for this gene. Introduction of disease causing mutations, identified in human CLC genes, abolished this capacity. Consistent with a similar function of both proteins, the green fluorescent protein-tagged AtCLC-d protein showed the identical localization pattern as the yeast ScCLC protein. This suggests that in Arabidopsis AtCLC-d functions as an intracellular chloride channel.

KW - Amino Acid Sequence

KW - Animals

KW - Arabidopsis/genetics

KW - Chloride Channels/chemistry

KW - Consensus Sequence

KW - DNA, Complementary/genetics

KW - DNA, Plant/chemistry

KW - Genes, Plant

KW - Genetic Complementation Test

KW - Humans

KW - Molecular Sequence Data

KW - Muscle Proteins/chemistry

KW - Polymerase Chain Reaction

KW - Transfection

KW - Xenopus laevis

U2 - 10.1074/jbc.271.52.33632

DO - 10.1074/jbc.271.52.33632

M3 - SCORING: Journal article

C2 - 8969232

VL - 271

SP - 33632

EP - 33638

JO - J BIOL CHEM

JF - J BIOL CHEM

SN - 0021-9258

IS - 52

ER -