title: Protein kinase C negatively regulates Akt activity and modifies UVC-induced apoptosis in mouse keratinocytes.
abstract: Skin keratinocytes are subject to frequent chemical and physical injury and have developed elaborate cell survival mechanisms to compensate. Among these, the Akt/protein kinase B (PKB) pathway protects keratinocytes from the toxic effects of ultraviolet light (UV). In contrast, the protein kinase C (PKC) family is involved in several keratinocyte death pathways. During an examination of potential interactions among these two pathways, we found that the insulin-like growth factor (IGF-1) activates both the PKC and the Akt signaling pathways in cultured primary mouse keratinocytes as indicated by increased phospho-PKC and phospho-Ser-473-Akt. IGF-1 also selectively induced translocation of PKCdelta and PKCepsilon from soluble to particulate fractions in mouse keratinocytes. Furthermore, the PKC-specific inhibitor, GF109203X, increased IGF-1-induced phospho-Ser-473-Akt and Akt kinase activity and enhanced IGF-1 protection from UVC-induced apoptosis. Selective activation of PKC by 12-O-tetradecanoylphorbol-13-acetate (TPA) reduced phospho-Ser-473-Akt, suggesting that activation of PKC inhibits Akt activity. TPA also attenuated IGF-1 and epidermal growth factor-induced phospho-Ser-473-Akt, reduced Akt kinase activity, and blocked IGF-1 protection from UVC-induced apoptosis. The inhibition of Akt activity by TPA was reduced by inhibitors of protein phosphatase 2A, and TPA stimulated the association of phosphatase 2A with Akt. Individual PKC isoforms were overexpressed in cultured keratinocytes by transduction with adenoviral vectors or inhibited with PKC-selective inhibitors. These studies indicated that PKCdelta and PKCepsilon were selectively potent at causing dephosphorylation of Akt and modifying cell survival, whereas PKCalpha enhanced phosphorylation of Akt on Ser-473. Our results suggested that activation of PKCdelta and PKCepsilon provide a negative regulation for Akt phosphorylation and kinase activity in mouse keratinocytes and serve as modulators of cell survival pathways in response to external stimuli.
authors.affiliation: Laboratory of Cellular Carcinogenesis and Tumor Promotion, Center for Cancer Research, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA. lilu@mail.nih.gov
authors.forename: Luowei
authors.initials: L
authors.lastname: Li
authors.forename: Keeran
authors.initials: K
authors.lastname: Sampat
authors.forename: Nancy
authors.initials: N
authors.lastname: Hu
authors.forename: Julia
authors.initials: J
authors.lastname: Zakari
authors.forename: Stuart H
authors.initials: SH
authors.lastname: Yuspa
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D015151:Immunoblotting; D047468:Immunoprecipitation; D007211:Indoles / Q000494:pharmacology; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D015603:Keratinocytes / Q000378:metabolism*; D008301:Maleimides / Q000494:pharmacology; D051379:Mice; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D010766:Phosphorylation; D020033:Protein Isoforms; D011493:Protein Kinase C / Q000378:metabolism*; D051745:Protein Kinase C-delta / Q000378:metabolism; D051744:Protein Kinase C-epsilon / Q000378:metabolism; D054648:Protein Phosphatase 2; D021381:Protein Transport; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D012694:Serine / Q000737:chemistry; D015398:Signal Transduction; D012867:Skin / Q000378:metabolism / Q000528:radiation effects; D013755:Tetradecanoylphorbol Acetate / Q000737:chemistry; D013997:Time Factors; D014466:Ultraviolet Rays
chemical_list: D007211:Indoles; D008301:Maleimides; D020033:Protein Isoforms; D012694:Serine; D007334:Insulin-Like Growth Factor I; D051057:Proto-Oncogene Proteins c-akt; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D051744:Protein Kinase C-epsilon; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2; C070515:bisindolylmaleimide I; D013755:Tetradecanoylphorbol Acetate