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Published 8 November 2004. doi:10.1083/jcb.200404142
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 167, Number 3, 555-562
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Article

 Protein kinase C spatially and temporally regulates gap junctional communication during human wound repair via phosphorylation of connexin43 on serine368

Theresa S. Richards1,2, Clarence A. Dunn1,2, William G. Carter1,2, Marcia L. Usui3, John E. Olerud3, and Paul D. Lampe1,2

1 Divisions of Basic and Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA 98109
2 Department of Pathobiology, Department of Medicine, University of Washington, Seattle, WA 98195
3 Division of Dermatology, Department of Medicine, University of Washington, Seattle, WA 98195

Correspondence to Paul D. Lampe: plampe{at}fhcrc.org

Phosphorylation of connexin43 (Cx43) on serine368 (S368) has been shown to decrease gap junctional communication via a reduction in unitary channel conductance. Examination of phosphoserine368 (pS368) in normal human skin tissue using a phosphorylation site–specific antibody showed relatively even distribution throughout the epidermal layers. However, 24 h after wounding, but not at 6 or 72 h, pS368 levels were dramatically increased in basal keratinocytes and essentially lost from suprabasal layers adjacent to the wound (i.e., within 200 µm of it). Scratch wounding of primary human keratinocytes caused a protein kinase C (PKC)-dependent increase in pS368 in cells adjacent to the scratch, with a time course similar to that found in the wounds. Keratinocytes at the edge of the scratch also transferred dye much less efficiently at 24 h, in a manner dependent on PKC. However, keratinocyte migration to fill the scratch required early (within <6 h) gap junctional communication. Our evidence indicates that PKC-dependent phosphorylation of Cx43 at S368 creates dynamic communication compartments that can temporally and spatially regulate wound healing.

T.S. Richards and C.A. Dunn contributed equally to this paper.

Abbreviations used in this paper: BIM, bisindolylmaleimide; CBX, carbenoxolone; Cx43, connexin43; Cx46, connexin46; Cx50, connexin50; HFK, human foreskin keratinocyte; pS368, phosphoserine368; S368, serine368.


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