Referenced in: none

Automatically associated channels: ClC4

Title: Altered gating and regulation of a carboxy-terminal ClC channel mutant expressed in the Caenorhabditis elegans oocyte.

Authors: Jerod Denton, Keith Nehrke, Xiaoyan Yin, Andrew M Beld, Kevin Strange

Journal, date & volume: Am. J. Physiol., Cell Physiol., 2006 Apr , 290, C1109-18

PubMed link: http://www.ncbi.nlm.nih.gov/pubmed/16306126

Abstract
CLH-3a and CLH-3b are swelling-activated, alternatively spliced Caenorhabditis elegans ClC anion channels that have identical membrane domains but exhibit marked differences in their cytoplasmic NH(2) and COOH termini. The major differences include a 71-amino acid CLH-3a NH(2)-terminal extension and a 270-amino acid extension of the CLH-3b COOH terminus. Splice variation gives rise to channels with striking differences in voltage, pH, and Cl(-) sensitivity. On the basis of structural and functional insights gained from crystal structures of bacterial ClCs, we suggested previously that these functional differences are due to alternative splicing of the COOH terminus that may change the accessibility and/or function of pore-associated ion-binding sites. We recently identified a mutant worm strain harboring a COOH-terminal deletion mutation in the clh-3 gene. This mutation removes 101 COOH-terminal amino acids unique to CLH-3b and an additional 64 upstream amino acids shared by both channels. CLH-3b is expressed in the worm oocyte, which allowed us to characterize the mutant channel, CLH-3bDeltaC, in its native cellular environment. CLH-3bDeltaC exhibits altered voltage-dependent gating as well as pH and Cl(-) sensitivity that resemble those of CLH-3a. This mutation also alters channel inhibition by Zn(2+), prevents ATP depletion-induced activation, and dramatically reduces volume sensitivity. These results suggest that the deleted COOH-terminal region of CLH-3bDeltaC functions to modulate channel sensitivity to voltage and extracellular ions. This region also likely plays a role in channel regulation and cell volume sensitivity. Our findings contribute to a growing body of evidence indicating that cytoplasmic domains play key roles in the gating and regulation of eukaryotic ClCs.