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PMID:16249326
| Citation |
Nakamura, A, Fukuda, A, Sakai, S and Tanaka, Y (2006) Molecular cloning, functional expression and subcellular localization of two putative vacuolar voltage-gated chloride channels in rice (Oryza sativa L.). Plant Cell Physiol. 47:32-42 |
|---|---|
| Abstract |
We isolated two cDNA clones (OsCLC-1 and OsCLC-2) homologous to tobacco CLC-Nt1, which encodes a voltage-gated chloride channel, from rice (Oryza sativa L. ssp. japonica, cv. Nipponbare). The deduced amino acid sequences were highly conserved (87.9% identity with each other). Southern blot analysis of the rice genomic DNA revealed that OsCLC-1 and OsCLC-2 were single-copy genes on chromosomes 4 and 2, respectively. OsCLC-1 was expressed in most tissues, whereas OsCLC-2 was expressed only in the roots, nodes, internodes and leaf sheaths. The level of expression of OsCLC-1, but not of OsCLC-2, was increased by treatment with NaCl. Both genes could partly substitute for GEF1, which encodes the sole chloride channel in yeast, by restoring growth under ionic stress. These results indicate that both genes are chloride channel genes. The proteins from both genes were immunochemically detected in the tonoplast fraction. Tagged synthetic green fluorescent protein which was fused to OsCLC-1 or OsCLC-2 localized in the vacuolar membranes. These results indicate that the proteins may play a role in the transport of chloride ions across the vacuolar membrane. We isolated loss-of-function mutants of both genes from a panel of rice mutants produced by the insertion of a retrotransposon, Tos17, in the exon region, and found inhibition of growth at all life stages. |
| Links |
PubMed Online version:10.1093/pcp/pci220 |
| Keywords |
Amino Acid Sequence; Base Sequence; Chloride Channels/genetics; Chloride Channels/metabolism; Chromosome Mapping; Cloning, Molecular; DNA, Complementary/genetics; DNA, Plant/genetics; Gene Expression/drug effects; Genes, Fungal; Genes, Plant; Molecular Sequence Data; Mutagenesis, Insertional; Oryza sativa/drug effects; Oryza sativa/genetics; Oryza sativa/metabolism; Plant Proteins/genetics; Plant Proteins/metabolism; Recombinant Proteins/genetics; Recombinant Proteins/metabolism; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae/metabolism; Saccharomyces cerevisiae Proteins/genetics; Saccharomyces cerevisiae Proteins/metabolism; Sequence Homology, Amino Acid; Sodium Chloride/pharmacology; Species Specificity; Subcellular Fractions/metabolism; Tissue Distribution |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0009705: plant-type vacuole membrane |
ECO:0000314: |
C |
Figure 7 |
complete | ||||
|
part_of |
GO:0009705: plant-type vacuole membrane |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
| GO:0009705: plant-type vacuole membrane |
ECO:0000314: |
C |
Figure 7 |
complete | ||||
|
part_of |
GO:0009705: plant-type vacuole membrane |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
See also
References
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