Casein kinase 1 regulates connexin-43 gap junction assembly

Cynthia D Cooper; Paul D Lampe

doi:10.1074/jbc.M209427200

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Casein kinase 1 regulates connexin-43 gap junction assembly

Journal article

Open access

Peer reviewed

Casein kinase 1 regulates connexin-43 gap junction assembly

Cynthia D Cooper and Paul D Lampe

The Journal of biological chemistry, Vol.277(47), pp.44962-44968

11/22/2002

DOI: https://doi.org/10.1074/jbc.M209427200

Handle:

https://hdl.handle.net/2376/114511

PMID: 12270943

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Abstract

Immunohistochemistry

Protein Kinases - metabolism

Gap Junctions - metabolism

Phosphorylation

Kidney - drug effects

Casein Kinases

Connexin 43 - metabolism

Cells, Cultured

Rats

Kidney - cytology

Recombinant Fusion Proteins - metabolism

Protein Kinase Inhibitors

Animals

Isoquinolines - pharmacology

Isoenzymes - metabolism

Phloroglucinol - pharmacology

Recombinant Fusion Proteins - genetics

Indoles - pharmacology

Cell Membrane - metabolism

Phloroglucinol - analogs & derivatives

Isoenzymes - antagonists & inhibitors

Connexin 43 - genetics

Phosphorylation of members of the connexin family of gap junction proteins has been correlated with gap junction assembly, but the mechanisms involved remain unclear. We have examined the role of casein kinase 1 (CK1) in connexin-43 (Cx43) gap junction assembly. Cellular co-immunoprecipitation experiments and in vitro CK1 phosphorylation reactions indicate that CK1 interacted with and phosphorylated Cx43, initially on serine(s) 325, 328, or 330. (32)P(i)-Metabolically labeled cells treated with CKI-7, a specific CK1 inhibitor, showed a reduction in Cx43 phosphorylation on site(s) that can be phosphorylated by CK1 in vitro. To examine CK1 function, normal rat kidney cells were treated with CKI-7, and Cx43 content was analyzed by Triton X-100 extraction, cell-surface biotinylation, and immunofluorescence. Western blot analysis indicated a slight increase in total Cx43, whereas gap junctional (Triton-insoluble) Cx43 decreased, and non-junctional plasma membrane Cx43 increased (as detected by cell surface biotinylation). Immunofluorescence experiments in the presence of CK1 inhibitor showed increases in Cx43 plasma membrane localization but not necessarily accumulation at cell-cell interfaces. Decreased gap junctional and phosphorylated Cx43 was also detected when cells were treated with IC261, a CK1 inhibitor specific for delta or epsilon isoforms. These data suggest CK1delta could regulate Cx43 gap junction assembly by directly phosphorylating Cx43.

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Title: Casein kinase 1 regulates connexin-43 gap junction assembly
Creators: Cynthia D Cooper - Fred Hutchinson Cancer Research Center and Department of Pathobiology, University of Washington, Seattle, Washington 98109-1024, USA
Paul D Lampe
Publication Details: The Journal of biological chemistry, Vol.277(47), pp.44962-44968
Academic Unit: Molecular Biosciences, School of
Publisher: United States
Grant note: R01 GM055632 / NIGMS NIH HHS GM55632 / NIGMS NIH HHS
Identifiers: 99900548291401842
Language: English
Resource Type: Journal article