title: v-Src phosphorylation of connexin 43 on Tyr247 and Tyr265 disrupts gap junctional communication.
abstract: The mechanism by which v-Src disrupts connexin (Cx)43 intercellular gap junctional communication (GJC) is not clear. In this study, we determined that Tyr247 (Y247) and the previously identified Tyr265 (Y265) site of Cx43 were the primary phosphorylation targets for activated Src in vitro. We established an in vivo experimental system by stably expressing v-Src and wild-type (wt) Cx43, or Y247F, Y265F, or Y247F/Y265F Cx43 mutants in a Cx43 knockout mouse cell line. Wt and mutant Cx43 localized to the plasma membrane in the absence or presence of v-Src. When coexpressed with v-Src, the Y247F, Y265F, and Y247F/Y265F Cx43 mutants exhibited significantly reduced levels of tyrosine phosphorylation compared with wt Cx43, indicating that Y247 and Y265 were phosphorylation targets of v-Src in vivo. Most importantly, GJC established by the Y247F, Y265F, and Y247F/Y265F Cx43 mutants was resistant to disruption by v-Src. Furthermore, we did not find evidence for a role for mitogen-activated protein kinase in mediating the disruption of GJC by v-Src. We conclude that phosphorylation on Y247 and Y265 of Cx43 is responsible for disrupting GJC in these mammalian cells expressing v-Src.
authors.affiliation: Molecular Carcinogenesis Section, Cancer Research Center of Hawaii, Honolulu, HI 96813, USA.
authors.forename: R
authors.initials: R
authors.lastname: Lin
authors.forename: B J
authors.initials: BJ
authors.lastname: Warn-Cramer
authors.forename: W E
authors.initials: WE
authors.lastname: Kurata
authors.forename: A F
authors.initials: AF
authors.lastname: Lau
mesh_terms: D002450:Cell Communication; D002462:Cell Membrane; D002999:Clone Cells; D018031:Connexin 43 / Q000235:genetics / Q000378:metabolism*; D017629:Gap Junctions / Q000502:physiology*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D008954:Models, Biological; D016297:Mutagenesis, Site-Directed; D015688:Oncogene Protein pp60(v-src) / Q000378:metabolism*; D010766:Phosphorylation; D012694:Serine; D014443:Tyrosine
chemical_list: D018031:Connexin 43; D014443:Tyrosine; D012694:Serine; D015688:Oncogene Protein pp60(v-src); D020928:Mitogen-Activated Protein Kinases