Conserved cytoplasmic tyrosine residues of the gamma subunit are required for a phagocytic signal mediated by Fc gamma RIIIA.
Fc receptors for immunoglobulins are found on many cells and are important in host defense. We transfected Fc gamma RIIIA, present on macrophages and natural killer (NK) cells, into COS-1 cells to study its role in phagocytosis and calcium mobilization in the absence of other Fc gamma receptors. Human Fc gamma RIIIA-alpha (CD16) was cotransfected with its associated chains, either Fc gamma RIIIA gamma or zeta. Both gamma and zeta were observed to induce a phagocytic signal, but gamma was at least sixfold more effective than zeta. Conservative substitution by phenylalanine of either one of the two cytoplasmic tyrosine residues in the gamma chain resulted in markedly diminished phagocytosis and calcium mobilization. Tyrphostin 23, an inhibitor of tyrosine kinases, reversibly inhibited phagocytosis. Further, in vitro kinase assays with the wild type and mutant gamma chains demonstrated that the wild type gamma chain, but not the mutant gamma chains, is phosphorylated. These results suggest that the cytoplasmic tyrosine residues and tyrosine phosphorylation are required for Fc gamma RIIIA to mediate two signal transduction events: phagocytosis and calcium mobilization.
Park
J G
JG
Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia 19104.
Murray
R K
RK
Chien
P
P
Darby
C
C
Schreiber
A D
AD
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D002396:Catechols / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D017124:Conserved Sequence; D003593:Cytoplasm / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009570:Nitriles / Q000494:pharmacology; D010587:Phagocytosis / Q000187:drug effects / Q000276:immunology*; D017354:Point Mutation; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D017452:Receptors, IgG / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000276:immunology*; D014162:Transfection; D014443:Tyrosine; D020032:Tyrphostins
D002396:Catechols; D009570:Nitriles; D017452:Receptors, IgG; D020032:Tyrphostins; C060152:tyrphostin 47; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D002118:Calcium