Calpain-induced downregulation of activated protein kinase C-alpha affects lung epithelial cell morphology.
A few minutes after mouse lung epithelial cell lines were treated with 12-O-tetradecanoylphorbol-13-acetate (TPA), the cells rounded up and pulled away from their neighbors. Several hours later, the cells flattened out to resume their original morphology. To begin to characterize the enzymology underlying these changes, the subcellular distribution and intracellular content of the TPA receptor, protein kinase C (PKC), and its putative endogenous regulator, the Ca(2+)-dependent protease, calpain, were investigated. Of eight PKC isozymes examined in several tumorigenic and nontumorigenic cell lines, all cells contained PKC-alpha, PKC-delta, and PKC-zeta. TPA rapidly (5 min) translocated PKC-alpha from the cytosol to the particulate fraction; PKC-alpha concentrations then decreased with continued TPA exposure. PKC-zeta levels and intracellular location were not affected. An inhibitor of PKC activity, GF 109203X, prevented the initial morphological change. The calpain II isozyme was also found in all cell lines, and its cellular content increased as a result of TPA treatment. Calpain inhibitor I did not affect the initial shape change but prevented subsequent flattening of the cells. We therefore conclude that PKC activation is required for the TPA-induced alterations in lung cell morphology and that calpain mediates the return to a flattened epithelial appearance.
Dwyer
L D
LD
Molecular Toxicology Program, School of Pharmacy, University of Colorado, Denver 80262.
Miller
A C
AC
Parks
A L
AL
Jaken
S
S
Malkinson
A M
AM
D000818:Animals; D002154:Calpain / Q000302:isolation & purification / Q000378:metabolism*; D002460:Cell Line; D002461:Cell Line, Transformed; D003600:Cytosol / Q000201:enzymology; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects / Q000201:enzymology; D007211:Indoles / Q000494:pharmacology; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008168:Lung / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D008301:Maleimides / Q000494:pharmacology; D051379:Mice; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D013997:Time Factors
D007211:Indoles; D007527:Isoenzymes; D008301:Maleimides; D011493:Protein Kinase C; D002154:Calpain; C070515:bisindolylmaleimide I; D013755:Tetradecanoylphorbol Acetate