CLIC6, a member of the intracellular chloride channel family, interacts with dopamine D(2)-like receptors.
To identify proteins interacting with the C-terminal cytoplasmic tail of the dopamine D(3) receptor (D(3)R), we used the two-hybrid system to screen a rat brain cDNA library. We isolated three partial cDNAs encoding, respectively, for the MUPP1 multi-PDZ protein, for the N-terminal region of radixin, for GIPC and for a ... 160-amino acid open reading frame sharing high homology with the human CLIC6, also identified as parchorin in rabbit. In the two-hybrid system, CLIC6 was also able to interact with the D(2)R and D(4)R. The interaction between D(3)R and CLIC6 was confirmed by the use of a GST-D(3)R C-terminus fusion protein and COS cell extracts transiently expressing epitope-tagged CLIC6. In adult rat brain, CLIC6 mRNA expression was restricted to the choroid plexus, the striatal proliferative subventricular zone and the cerebellum where it is co-expressed with the D(3)R in the Purkinje cells of the lobules IX and X. CLIC6 mRNA was also detected in the pituitary in the posterior lobe and in cells co-expressing the D(2)R at the border between the intermediate and anterior lobes. In transfected HEK293 cells, D(3)R and CLIC6 co-localized at the plasma-membrane. No effect of CLIC6 transfection was observed on either intracellular chloride concentration or D(3)R/D(2)R-mediated response. In two-hybrid system, CLIC6 also interacted with MUPP1 and radixin but not GIPC, suggesting it could take part in a complex with D(2)-like receptors, not only by direct interaction with their C-termini, but also through interactions with scaffolding proteins.
Mesh Terms:
Amino Acid Sequence, Animals, Blotting, Northern, Brain, CHO Cells, COS Cells, Carrier Proteins, Cell Membrane, Cercopithecus aethiops, Chloride Channels, Chlorides, Choroid Plexus, Cloning, Molecular, Cricetinae, Extracellular Space, Humans, In Situ Hybridization, Molecular Sequence Data, Nerve Tissue Proteins, Neuropeptides, Peptide Fragments, Pituitary Gland, Protein Binding, RNA, Messenger, Rats, Receptors, Dopamine D2, Receptors, Dopamine D3, Thymidine, Transfection, Two-Hybrid System Techniques, Yeasts
Amino Acid Sequence, Animals, Blotting, Northern, Brain, CHO Cells, COS Cells, Carrier Proteins, Cell Membrane, Cercopithecus aethiops, Chloride Channels, Chlorides, Choroid Plexus, Cloning, Molecular, Cricetinae, Extracellular Space, Humans, In Situ Hybridization, Molecular Sequence Data, Nerve Tissue Proteins, Neuropeptides, Peptide Fragments, Pituitary Gland, Protein Binding, RNA, Messenger, Rats, Receptors, Dopamine D2, Receptors, Dopamine D3, Thymidine, Transfection, Two-Hybrid System Techniques, Yeasts
Brain Res. Mol. Brain Res.
Date: Sep. 10, 2003
PubMed ID: 14499480
View in: Pubmed Google Scholar
Download Curated Data For This Publication
2743
Switch View:
- Interactions 7