Splice variants of a ClC-2 chloride channel with differing functional characteristics |
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Authors: | Cid L P Niemeyer M I Ramírez A Sepúlveda F V |
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Affiliation: | Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago-7, Chile. pcid@cecs.cl |
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Abstract: | We identified two ClC-2 clones in a guinea pigintestinal epithelial cDNA library, one of which carries a 30-bpdeletion in the NH2 terminus. PCR using primersencompassing the deletion gave two products that furthermore wereamplified with specific primers confirming their authenticity. Thecorresponding genomic DNA sequence gave a structure of three exons andtwo introns. An internal donor site occurring within one of the exonsaccounts for the deletion, consistent with alternative splicing.Expression of the variants gpClC-2 and gpClC-277-86 in HEK-293cells generated inwardly rectifying chloride currents with similaractivation characteristics. Deactivation, however, occurred with fasterkinetics in gpClC-277-86. Site-directed mutagenesis suggeststhat a protein kinase C-mediated phosphorylation consensus site lost ingpClC-277-86 is not responsible for the observed change. Thedeletion-carrying variant is found in most tissues examined, and itappears more abundant in proximal colon, kidney, and testis. Thepresence of a splice variant of ClC-2 modified in itsNH2-terminal domain could have functional consequences intissues where their relative expression levels are different. |
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