title: Autophosphorylation of Dictyostelium myosin II heavy chain-specific protein kinase C is required for its activation and membrane dissociation.
abstract: Myosin II heavy chain (MHC)-specific protein kinase C (MHC-PKC) isolated from the ameba, Dictyostelium discoideum, regulates myosin II assembly and localization in response to the chemoattractant cAMP. cAMP stimulation of Dictyostelium cells leads to translocation of MHC-PKC from the cytosol to the membrane fraction, as well as causing an increase in both MHC-PKC phosphorylation and its kinase activity. MHC-PKC undergoes autophosphorylation with each mole of kinase incorporating about 20 mol of phosphate. The MHC-PKC autophosphorylation sites are thought to be located within a domain at the COOH-terminal region of MHC-PKC that contains a cluster of 21 serine and threonine residues. Here we report that deletion of this domain abolished the ability of the enzyme to undergo autophosphorylation in vitro. Furthermore, after this deletion, cAMP-dependent autophosphorylation of MHC-PKC as well as cAMP-dependent increases in kinase activity and subcellular localization were also abolished. These results provide evidence for the role of autophosphorylation in the regulation of MHC-PKC and indicate that this MHC-PKC autophosphorylation is required for the kinase activation in response to cAMP and for subcellular localization.
authors.affiliation: Department of Biochemistry, Hadassah Medical School, The Hebrew University, Jerusalem, Israel.
authors.forename: A
authors.initials: A
authors.lastname: Dembinsky
authors.forename: H
authors.initials: H
authors.lastname: Rubin
authors.forename: S
authors.initials: S
authors.lastname: Ravid
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D000242:Cyclic AMP / Q000378:metabolism; D004023:Dictyostelium; D004789:Enzyme Activation; D018995:Myosin Heavy Chains / Q000378:metabolism*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*
chemical_list: D000242:Cyclic AMP; D011493:Protein Kinase C; D018995:Myosin Heavy Chains