title: Double-stranded RNA-dependent protein kinase (PKR) is downregulated by phorbol ester.
abstract: The double-stranded RNA-dependent protein kinase (PKR) is one of the key mediators of interferon (IFN) action against certain viruses. PKR also plays an important role in signal transduction and immunomodulation. Understanding the regulation of PKR activity is important for the use of PKR as a tool to discover and develop novel therapeutics for viral infections, cancer and immune dysfunction. We found that phorbol 12-myristate 13-acetate (PMA), a potent activator of protein kinase C (PKC), decreased the level of autophosphorylated PKR in a dose- and time-dependent manner in IFN-treated mouse fibroblast cells. Polyinosinic-polycytidylic acid (poly I:C) treatment enhanced the activity of PKR induced by IFN, but did not overcome the PMA-induced reduction of PKR autophosphorylation. Western blot analysis with a monoclonal antibody to mouse PKR revealed that the decrease of PKR autophosphorylation in cells by PMA was a result of PKR protein degradation. Selective PKC inhibitors blocked the degradation of PKR stimulated by PMA, indicating that PKC activity was required for the effect. Furthermore, we also found that proteasome inhibitors prevented PMA-induced down regulation of PKR, indicating that an active proteasome is required. Our results identify a novel mechanism for the post-translational regulation of PKR.
authors.affiliation: Department of Chemistry, Cleveland State University, OH 44115, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhou
authors.forename: Barbara I
authors.initials: BI
authors.lastname: Chase
authors.forename: Mark
authors.initials: M
authors.lastname: Whitmore
authors.forename: Bryan R G
authors.initials: BR
authors.lastname: Williams
authors.forename: Aimin
authors.initials: A
authors.lastname: Zhou
mesh_terms: D000818:Animals; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects; D051379:Mice; D041681:NIH 3T3 Cells; D010703:Phorbol Esters / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D012330:RNA, Double-Stranded / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D019892:eIF-2 Kinase / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D010703:Phorbol Esters; D012330:RNA, Double-Stranded; D019892:eIF-2 Kinase; D011493:Protein Kinase C