Molecular cloning and characterization of a mitogen-activated protein kinase-associated intracellular chloride channel

J Biol Chem. 1999 Jan 15;274(3):1621-7. doi: 10.1074/jbc.274.3.1621.

Abstract

ERK7, a member of the mitogen-activated protein kinase family, has a carboxyl-terminal tail that is required for ERK7 activation, cellular localization, and its ability to inhibit DNA synthesis. To identify proteins that interact with ERK7, we utilized a yeast two-hybrid screen with the COOH-terminal tail of ERK7 as bait and isolated the cDNA for a novel protein termed CLIC3. The interaction between CLIC3 and ERK7 in mammalian cells was confirmed by co-immunoprecipitation. CLIC3 has significant homology to human intracellular chloride channels 1 (NCC27/CLIC1) and 2 and bovine kidney chloride channel p64. Like NCC27/CLIC1, CLIC3 is predominantly localized in the nucleus and stimulates chloride conductance when expressed in cells. Taken together, these results suggest that CLIC3 is a new member of the human CLIC family. The observed interaction between CLIC3 and ERK7 is the first demonstration of a stable complex between a protein that activates chloride ion transport and a member of the mitogen-activated protein kinase family of signal transducers. The specific association of CLIC3 with the COOH-terminal tail of ERK7 suggests that CLIC3 may play a role in the regulation of cell growth.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Northern
  • COS Cells
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism*
  • Cattle
  • Cell Nucleus / chemistry
  • Cell Nucleus / metabolism
  • Cells, Cultured
  • Chloride Channels / chemistry
  • Chloride Channels / genetics
  • Chloride Channels / metabolism*
  • Chloride Channels / physiology
  • Cloning, Molecular
  • Electrophysiology
  • Enzyme Activation
  • Humans
  • Kidney / chemistry
  • Kidney / metabolism
  • Molecular Sequence Data
  • Sequence Alignment

Substances

  • CLIC3 protein, human
  • Chloride Channels
  • Calcium-Calmodulin-Dependent Protein Kinases

Associated data

  • GENBANK/AF102166