title: ARF1-regulated coatomer directs the steady-state localization of protein kinase C epsilon at the Golgi apparatus.
abstract: Protein kinase C epsilon (PKCε) contributes to multiple signaling pathways affecting human disease. The function of PKCε requires it to undergo changes in subcellular distribution in response to signaling events. While the mechanisms underlying this translocation are incompletely understood, it involves the receptor for activated C kinase protein (RACK2/β'-COP). This receptor also functions as a vesicle coat protein in the secretory pathway where it is regulated by the small GTP-binding protein ADP-ribosylation factor, ARF1. We inhibited ARF1 activation to test the requirement for RACK2/β'-COP in PKCε localization in NIH3T3 fibroblasts. We found that steady-state localization of PKCε at the Golgi complex requires ARF1-regulated RACK2/β'-COP function. By contrast, we did not observe any defects in phorbol ester-induced translocation when ARF1 was inhibited. We also found that PKCε bound to isolated membranes through two distinct mechanisms. One mechanism was dependent upon RACK2/β'-COP while a second was RACK2/β'-COP-independent and stimulated by phorbol esters. Finally, we show that RACK2/β'-COP affects the subcellular distribution of a constitutively active form of PKCε, in a manner similar to what we observed for wild-type PKCε. Together, our data support a role for RACK2/β'-COP in the steady-state localization of PKCε at the Golgi apparatus, which may be independent of its role during PKCε translocation to the cell surface.
authors.affiliation: Department of Molecular Physiology & Biophysics, Roy J. and Lucille A. Carver College of Medicine, The University of Iowa, Iowa City, IA 52242, USA.
authors.forename: Tabitha A
authors.initials: TA
authors.lastname: Peterson
authors.forename: Mark
authors.initials: M
authors.lastname: Stamnes
mesh_terms: D020823:ADP-Ribosylation Factor 1 / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D022181:COP-Coated Vesicles / Q000187:drug effects / Q000378:metabolism*; D002417:Cattle; D020756:Coatomer Protein / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D010703:Phorbol Esters / Q000494:pharmacology; D051744:Protein Kinase C-epsilon / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D020756:Coatomer Protein; D010703:Phorbol Esters; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface; D051744:Protein Kinase C-epsilon; D020823:ADP-Ribosylation Factor 1