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v-Src-mediated phosphorylation of connexin43 on tyrosine disrupts gap junctional communication in mammalian cells

R Lin1, B J Warn-Cramer, W E Kurata

  • 1Cancer Research Center of Hawaii and the Department of Cell and Molecular Biology, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu 96813, USA. rui@hawaii.edu

Cell Communication & Adhesion
|June 18, 2002
PubMed

Insights

v-Src oncoprotein disrupts connexin43 (Cx43) gap junctional communication (GJC) by phosphorylating Cx43 at Y265 and Y247. Mutants lacking these sites resist v-Src-induced GJC disruption, revealing a phosphorylation-dependent mechanism.

Area of Science:

  • Cellular Biology
  • Oncology
  • Biochemistry

Background:

  • The v-Src oncoprotein is known to disrupt gap junctional communication (GJC) mediated by connexin43 (Cx43) in mammalian cells.
  • The precise molecular mechanisms by which v-Src interferes with Cx43 function remain incompletely understood.

Purpose of the Study:

  • To investigate the role of specific tyrosine phosphorylation sites (Y247 and Y265) on Cx43 in mediating v-Src-induced disruption of GJC.
  • To elucidate the sequential phosphorylation events on Cx43 by v-Src in vivo.

Main Methods:

  • Established a stable expression system in a Cx43 knockout mouse cell line co-expressing v-Src with wild-type (wt) Cx43 or Cx43 mutants (Y247F, Y265F, Y247F/Y265F).
  • Quantified phosphotyrosine (pTyr) levels of Cx43 and its mutants in the presence of v-Src.
  • Measured GJC function using the established cell system.

Main Results:

  • v-Src significantly reduced phosphotyrosine levels at Y247 and Y265 in Cx43 mutants compared to wt Cx43.
  • Phosphorylation at Y265 was necessary for efficient phosphorylation at Y247.
  • Cx43 mutants at Y247F, Y265F, and Y247F/Y265F exhibited resistance to v-Src-mediated disruption of GJC.

Conclusions:

  • Tyrosine phosphorylation of Cx43 at Y265, followed by Y247, is a critical mechanism by which v-Src disrupts GJC.
  • Cx43 phosphorylation sites Y247 and Y265 are direct in vivo targets of v-Src.
  • Understanding this processive phosphorylation pathway offers insights into cancer-related communication disruptions.

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