Differential role of actin, clathrin, and dynamin in Fc gamma receptor-mediated endocytosis and phagocytosis.
Clustering of macrophage Fc gamma receptors by multimeric immunoglobulin complexes leads to their internalization. Formation of small aggregates leads to endocytosis, whereas large particulate complexes induce phagocytosis. In RAW-264.7 macrophages, Fc gamma receptor endocytosis was found to be dependent on clathrin and dynamin and insensitive to cytochalasin. Clathrin also associates with nascent phagosomes, and earlier observations suggested that it plays an essential role in phagosome formation. However, we find that phagocytosis of IgG-coated large (> or =3 microm) particles was unaffected by inhibition of dynamin or by reducing the expression of clathrin using antisense mRNA but was eliminated by cytochalasin, implying a distinct mechanism dependent on actin assembly. The uptake of smaller particles (< or =1 microm) was only partially blocked by cytochalasin. Remarkably, the cytochalasin-resistant component was also insensitive to dominant-negative dynamin I and to clathrin antisense mRNA, implying the existence of a third internalization mechanism, independent of actin, dynamin, and clathrin. The uptake of small particles occurred by a process distinct from fluid phase pinocytosis, because it was not inhibited by dominant-negative Rab5. The insensitivity of phagocytosis to dominant-negative dynamin I enabled us to test the role of dynamin in phagosomal maturation. Although internalization of receptors from the plasma membrane was virtually eliminated by the K44A and S45N mutants of dynamin I, clearance of transferrin receptors and of CD18 from maturing phagosomes was unaffected by these mutants. This implies that removal of receptors from the phagosomal membrane occurs by a mechanism that is different from the one mediating internalization of the same receptors at the plasma membrane. These results imply that, contrary to prevailing notions, normal dynamin and clathrin function is not required for phagocytosis and reveal the existence of a component of phagocytosis that is independent of actin and Rab5.
Tse
Shirley M L
SM
Department of Pediatrics, Division of Rheumatology, Hospital for Sick Children, Institute of Medical Science, and University Health Network, University of Toronto, Toronto, Ontario M5G 1X8, Canada.
Furuya
Wendy
W
Gold
Elizabeth
E
Schreiber
Alan D
AD
Sandvig
Kirsten
K
Inman
Robert D
RD
Grinstein
Sergio
S
D000199:Actins / Q000235:genetics / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology; D002966:Clathrin / Q000235:genetics / Q000502:physiology*; D034284:Dynamin I / Q000235:genetics / Q000502:physiology*; D004705:Endocytosis / Q000276:immunology / Q000502:physiology*; D005786:Gene Expression Regulation / Q000276:immunology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D007074:Immunoglobulin G; D008164:Luminescent Proteins / Q000235:genetics; D008264:Macrophages / Q000276:immunology; D051379:Mice; D010587:Phagocytosis / Q000276:immunology / Q000502:physiology*; D010588:Phagosomes / Q000378:metabolism; D021381:Protein Transport; D016372:RNA, Antisense; D012333:RNA, Messenger / Q000235:genetics; D017452:Receptors, IgG / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic / Q000276:immunology
D000199:Actins; D002966:Clathrin; D007074:Immunoglobulin G; D008164:Luminescent Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D017452:Receptors, IgG; D049452:Green Fluorescent Proteins; D034284:Dynamin I