Dissection of the molecular basis of pp60(v-src) induced gating of connexin 43 gap junction channels

Lan Zhou, Eileen M. Kasperek, Bruce J. Nicholson

Research output: Contribution to journalArticlepeer-review

140 Scopus citations

Abstract

Suppression of gap-junctional communication by various protein kinases, growth factors, and oncogenes frequently correlates with enhanced mitogenesis. The oncogene v-src appears to cause acute closure of gap junction channels. Tyr265 in the COOH-terminal tail of connexin 43 (Cx43) has been implicated as a potential target of v-src, although v-src action has also been associated with changes in serine phosphorylation. We have investigated the mechanism of this acute regulation through mutagenesis of Cx43 expressed in Xenopus laevis oocyte pairs. Truncations of the COOH- terminal domain led to an almost complete loss of response of Cx43 to v-src, but this was restored by coexpression of the independent COOH-terminal polypeptide. This suggests a ball and chain gating mechanism, similar to the mechanism proposed for pH gating of Cx43, and K+ channel inactivation. Surprisingly, we found that v-src mediated gating of Cx43 did not require the tyrosine site, but did seem to depend on the presence of two potential SH3 binding domains and the mitogen-activated protein (MAP) kinase phosphorylation sites within them. Further point mutagenesis and pharmacological studies in normal rat kidney (NRK) cells implicated MAP kinase in the gating response to v-src, while the stable binding of v-src to Cx43 (in part mediated by SH3 domains) did not correlate with its ability to mediate channel closure. This suggests a common link between closure of gap junctions by v-src and other mitogens, such as EGF and lysophosphatidic acid (LPA).

Original languageEnglish (US)
Pages (from-to)1033-1045
Number of pages13
JournalJournal of Cell Biology
Volume144
Issue number5
DOIs
StatePublished - Mar 8 1999

Keywords

  • Intercellular coupling
  • MAP kinase
  • Phosphorylation
  • V-src
  • Xenopus oocytes

ASJC Scopus subject areas

  • Cell Biology

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