The alpha isoform of protein kinase C is involved in signaling the response of desmosomes to wounding in cultured epithelial cells.
Initiation of reepithelialization upon wounding is still poorly understood. To enhance this understanding, we focus here on changes in the adhesive state of desmosomes of cultured Madin-Darby canine kidney cells in response to wounding of confluent cell sheets. Previous results show that desmosomal adhesion in Madin-Darby canine kidney cells changes from a calcium-dependent state to calcium independence in confluent cell sheets. We show that this change, which requires culture confluence to develop, is rapidly reversed upon wounding of confluent cell sheets. Moreover, the change to calcium dependence in wound edge cells is propagated to cells hundreds of micrometers away from the wound edge. Rapid transition from calcium independence to calcium dependence also occurs when cells are treated with phorbol esters that activate PKC. PKC inhibitors, including the conventional isoform inhibitor Gö6976, cause rapid transition from calcium dependence to calcium independence, even in subconfluent cells. The cellular location of the alpha isoform of PKC correlates with the calcium dependence of desmosomes. Upon monolayer wounding, PKCalpha translocates rapidly to the cell periphery, becomes Triton X-100 insoluble, and also becomes concentrated in lamellipodia. The PKCalpha translocation upon wounding precedes both the increase in PKC activity in the membrane fraction and the reversion of desmosomes to calcium dependence. Specific depletion of PKCalpha with an antisense oligonucleotide increases the number of cells with calcium-independent desmosomes. These results show that PKCalpha participates in a novel signaling pathway that modulates desmosomal adhesion in response to wounding.
Wallis
S
S
School of Biological Sciences, University of Manchester, Manchester, M13 9PT, United Kingdom.
Lloyd
S
S
Wise
I
I
Ireland
G
G
Fleming
T P
TP
Garrod
D
D
D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002118:Calcium / Q000378:metabolism; D002452:Cell Count; D002465:Cell Movement; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000032:analysis; D003896:Desmosomes / Q000502:physiology*; D004285:Dogs; D004847:Epithelial Cells / Q000502:physiology; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D011493:Protein Kinase C / Q000235:genetics / Q000502:physiology*; D051571:Protein Kinase C-alpha; D015398:Signal Transduction / Q000502:physiology; D013997:Time Factors; D014945:Wound Healing / Q000502:physiology*
D018076:DNA, Complementary; D007527:Isoenzymes; C496628:PRKCA protein, human; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D002118:Calcium