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PMID: 8241569 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Epidermal growth factor stimulates the disruption of gap junctional communication and connexin43 phosphorylation independent of 12-0-tetradecanoylphorbol 13-acetate-sensitive protein kinase C: the possible involvement of mitogen-activated protein kinase.

Molecular biology of the cell ·Vol. 4 ·No. 8 ·1993-08-00 ·Pages 837-48

Kanemitsu MY, Lau AF

Abstract

We previously reported that epidermal growth factor (EGF) induced the disruption of gap junctional communication (gjc) and serine phosphorylation of connexin43 (Cx43) in T51B rat liver epithelial cells. However, the cascade of events linking EGF receptor activation to these particular responses have not been fully characterized. Furthermore, the serine kinase(s) acting directly on Cx43 remain unidentified. In the current study, we demonstrate that downmodulation of 12-0-tetradecanoylphorbol 13-acetate (TPA)-sensitive protein kinase C (PKC) activity does not affect EGF's ability to reduce junctional permeability or phosphorylate Cx43 in T51B cells. EGF in the presence or absence of chronic TPA treatment stimulated marked increases in Cx43 phosphorylation on numerous sites as determined by two-dimensional tryptic phosphopeptide mapping. Computer-assisted sequence analysis of Cx43 identified several protein kinase phosphorylation consensus sites including two sites for mitogen-activated protein (MAP) kinase. EGF stimulated activation of MAP kinase in a time- and dose-dependent manner where the kinetics of kinase activity corroborated its possible involvement in mediating EGF's effects. Moreover, purified MAP kinase directly phosphorylated Cx43 on serine residues in vitro. Two-dimensional tryptic and chymotryptic phosphopeptide mapping demonstrated that the in vitro phosphopeptides represented a specific subset of the in vivo phosphopeptides produced in response to EGF after chronic TPA treatment. Therefore, EGF-induced disruption of gjc and phosphorylation of Cx43 may be mediated in part by MAP kinase in vivo.

MeSH Terms
Animals Cell Communication Cell Line Connexin 43/metabolism Down-Regulation Electrophoresis, Gel, Two-Dimensional Epidermal Growth Factor/metabolism Gap Junctions Mitogen-Activated Protein Kinase 1 Phosphorylation Protein Kinase C/drug effects,metabolism Protein Serine-Threonine Kinases/metabolism Protein-Tyrosine Kinases/metabolism Rats Tetradecanoylphorbol Acetate/pharmacology
Chemicals
Connexin 43 Epidermal Growth Factor Protein-Tyrosine Kinases Protein Serine-Threonine Kinases Protein Kinase C Mitogen-Activated Protein Kinase 1 Tetradecanoylphorbol Acetate
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kanemitsu M Y
Molecular Carcinogenesis Program, Cancer Research Center of Hawaii, Honolulu.
Lau A F
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.4.8.837
Published
1993-08-00
Pages
837-48
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC300996
Subset
IM
Grants
NCI NIH HHS · CA52098 · United States
NCRR NIH HHS · RCMI RR030601 · United States
External Links
PubMed source DOI: 10.1091/mbc.4.8.837 PMC Full Text View journal details
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