Protein kinase C-alpha activity modulates transepithelial permeability and cell junctions in the LLC-PK1 epithelial cell line.
Modulation of protein kinase C (PKC) by 12-O-tetradecanoylphorbol-13-acetate (TPA) disrupts the cell-cell junctions of the epithelial cell line LLC-PK1. To examine the role of specific PKC isoforms in this process we have created modified LLC-PK1 subclones that express wild-type and dominant negative versions of PKC-alpha under control of the tetracycline-responsive expression system. Overexpression of wild-type PKC-alpha rendered the cells more sensitive to the effects of TPA on transepithelial permeability as measured by loss of transepithelial resistance across the cell sheet. Conversely, expression of a dominant negative PKC-alpha rendered the cells more resistant to the effects of TPA as measured both by loss of transepithelial resistance as well as cell scattering. The properties of both subclones could be modulated by the addition of tetracycline, which suppressed the effect of the exogenous genes. These results indicate that the alpha isoform of PKC is at least one of the isoforms that regulate tight junctions and other cell-cell junctions of LLC-PK1 epithelia.
Rosson
D
D
Lankenau Medical Research Center, Wynnewood, Pennsylvania 19096-3411, USA.
O'Brien
T G
TG
Kampherstein
J A
JA
Szallasi
Z
Z
Bogi
K
K
Blumberg
P M
PM
Mullin
J M
JM
D000409:Alanine; D000818:Animals; D001665:Binding Sites; D002463:Cell Membrane Permeability / Q000187:drug effects; D002999:Clone Cells; D007365:Intercellular Junctions / Q000502:physiology* / Q000648:ultrastructure; D007527:Isoenzymes / Q000096:biosynthesis / Q000378:metabolism*; D007700:Kinetics; D018374:LLC-PK1 Cells; D008239:Lysine; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D011493:Protein Kinase C / Q000096:biosynthesis / Q000378:metabolism*; D051571:Protein Kinase C-alpha; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D013552:Swine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection
D007527:Isoenzymes; D011994:Recombinant Proteins; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D008239:Lysine; D013755:Tetradecanoylphorbol Acetate; D000409:Alanine