title: Phosphorylation of FcgammaRIIA is required for the receptor-induced actin rearrangement and capping: the role of membrane rafts.
abstract: Activation of Fcgamma receptor II (FcgammaRII) induces rearrangement of the actin-based cytoskeleton that serves as a driving force for FcgammaRII-mediated phagocytosis and FcgammaRII capping. To get insight into the signaling events that lead to the actin reorganization we investigated the role of raft-associated Src family tyrosine kinases in capping of FcgammaRII in U937 cells. After crosslinking, FcgammaRII was found to be recruited to detergent-resistant membrane domains (DRMs), rafts, where it coexisted with Lyn kinase and underwent tyrosine phosphorylation. Lyn was displaced from DRMs under the influence of DL-alpha-hydroxymyristic acid and 2-bromopalmitic acid, agents blocking N-terminal myristoylation and palmitoylation of proteins, respectively, and after disruption of DRM integrity by depletion of plasma membrane cholesterol with beta-cyclodextrin. Under these conditions, phosphorylation of the crosslinked FcgammaRII was diminished and assembly of FcgammaRII caps was blocked. The similar reduction of FcgammaRII cap formation correlated with inhibition of receptor phosphorylation was achieved with the use of PP1 and herbimycin A, specific inhibitors of Src family tyrosine kinases. Phosphorylation of FcgammaRIIA expressed in BHK cells, lacking endogenous FcgammaRs, was abolished by substitution of tyrosine 298 by phenylalanine in the ITAM of the receptor. The mutant receptor did not undergo translocation towards cap-like structures and failed to promote the receptor-mediated spreading of the cells, as compared to BHK cells transfected with the wild-type FcgammaRIIA. On the basis of these data, we suggest that tyrosine phosphorylation of activated FcgammaRIIA by raft-residing tyrosine kinases of the Src family triggers signaling pathways that control the rearrangement of the actin cytoskeleton required for FcgammaRII-mediated motility.
authors.affiliation: Department of Cell Biology, Nencki Institute of Experimental Biology, 3 Pasteur St., 02-093 Warsaw, Poland. Universität Bielefeld, Fakultät für Chemie, Biochemie II, 33615 Bielefeld, Germany.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Kwiatkowska
authors.forename: Jürgen
authors.initials: J
authors.lastname: Frey
authors.forename: Andrzej
authors.initials: A
authors.lastname: Sobota
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000818:Animals; D015703:Antigens, CD / Q000187:drug effects / Q000378:metabolism*; D002460:Cell Line; D002784:Cholesterol / Q000172:deficiency; D006224:Cricetinae; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005057:Eukaryotic Cells / Q000187:drug effects / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006801:Humans; D021962:Membrane Microdomains / Q000187:drug effects / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D010168:Palmitates / Q000494:pharmacology; D010587:Phagocytosis / Q000187:drug effects / Q000502:physiology; D010766:Phosphorylation / Q000187:drug effects; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism*; D017452:Receptors, IgG / Q000187:drug effects / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D004791:Enzyme Inhibitors; C080568:Fc gamma receptor IIA; D010168:Palmitates; D011956:Receptors, Cell Surface; D017452:Receptors, IgG; C022776:2-bromopalmitate; D014443:Tyrosine; D002784:Cholesterol; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases