Protein kinase C spatially and temporally regulates gap junctional communication during human wound repair via phosphorylation of connexin43 on serine368.
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 microm 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.
Richards
Theresa S
TS
Division of Basic and Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
Dunn
Clarence A
CA
Carter
William G
WG
Usui
Marcia L
ML
Olerud
John E
JE
Lampe
Paul D
PD
D002450:Cell Communication; D002465:Cell Movement; D018031:Connexin 43 / Q000378:metabolism*; D017629:Gap Junctions / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D015603:Keratinocytes / Q000201:enzymology / Q000473:pathology / Q000502:physiology; D007700:Kinetics; D010766:Phosphorylation; D010768:Phosphoserine / Q000032:analysis; D011493:Protein Kinase C / Q000378:metabolism / Q000502:physiology*; D012867:Skin / Q000737:chemistry / Q000166:cytology; D014945:Wound Healing
D018031:Connexin 43; D010768:Phosphoserine; D011493:Protein Kinase C