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J. Cell Biol., Volume 143, Number 6, December 14, 1998 1735-1747

Cellular Interaction of Integrin alpha 3beta 1 with Laminin 5 Promotes Gap Junctional Communication

Paul D. Lampe,*Dagger Beth P. Nguyen,*Dagger Susana Gil,* Marcia Usui,§ John Olerud,§ Yoshikazu Takada,parallel and William G. Carter*Dagger

* Divisions of Basic Sciences and Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109; Dagger  Department of Pathobiology and § Division of Dermatology, University of Washington, Seattle, Washington 98105; and parallel  Department of Vascular Biology, The Scripps Research Institute, La Jolla, California 92037

Wounding of skin activates epidermal cell migration over exposed dermal collagen and fibronectin and over laminin 5 secreted into the provisional basement membrane. Gap junctional intercellular communication (GJIC) has been proposed to integrate the individual motile cells into a synchronized colony. We found that outgrowths of human keratinocytes in wounds or epibole cultures display parallel changes in the expression of laminin 5, integrin alpha 3beta 1, E-cadherin, and the gap junctional protein connexin 43. Adhesion of keratinocytes on laminin 5, collagen, and fibronectin was found to differentially regulate GJIC. When keratinocytes were adhered on laminin 5, both structural (assembly of connexin 43 in gap junctions) and functional (dye transfer) assays showed a two- to threefold increase compared with collagen and five- to eightfold over fibronectin. Based on studies with immobilized integrin antibody and integrin-transfected Chinese hamster ovary cells, the interaction of integrin alpha 3beta 1 with laminin 5 was sufficient to promote GJIC. Mapping of intermediate steps in the pathway linking alpha 3beta 1-laminin 5 interactions to GJIC indicated that protein trafficking and Rho signaling were both required. We suggest that adhesion of epithelial cells to laminin 5 in the basement membrane via alpha 3beta 1 promotes GJIC that integrates individual cells into synchronized epiboles.

Key words: gap junctionsintegrinslaminin 5epidermisintracellular protein trafficking


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