Endocytosis of FcalphaR is clathrin and dynamin dependent, but its cytoplasmic domain is not required.
FcalphaR, the Fc receptor for IgA, is essential for IgA-mediated immune responses. Previous studies have shown that IgA and IgA immune complexes can be rapidly endocytosed by FcalphaR. However, the underlying mechanism remains unclear. Here, we investigated the endocytic pathway of FcalphaR in monocytic cell line, U937, that naturally express FcalphaR and in transfected Chinese hamster ovary (CHO), COS-7 and Hela cells. By using selective chemical inhibitors of different endocytic pathways, overexpression of dominant-negative mutants of Eps15 and knockdown of clathrin heavy chain (CHC) via RNA interference, we demonstrated that endocytosis of FcalphaR was through a clathrin-mediated pathway. The endocytosed FcalphaR went into Rab5- and Rab11-positive endosomes. However, endocytosis of FcalphaR could not be blocked by a dominant-negative mutant of Rab5. We also demonstrated that endocytosis of FcalphaR was dynamin-dependent by overexpressing a dominant-negative mutant of dynamin. The potential endocytic motif for FcalphaR was also examined. Unexpectedly, we found that the entire cytoplasmic domain of FcalphaR was not required for the endocytic process of FcalphaR. We conclude that endocytosis of FcalphaR is clathrin- and dynamin-dependent, but is not regulated by Rab5, and the endocytic motif is not located in the cytoplasmic domain of FcalphaR.
Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing 100005, China.
Min
M
Peng
Na
N
Yin
Wei
W
Zhang
D000818:Animals; D015703:Antigens, CD / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D016466:CHO Cells; D019556:COS Cells; D002522:Chlorocebus aethiops; D002966:Clathrin / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006224:Cricetinae; D003412:Cricetulus; D003593:Cytoplasm / Q000378:metabolism; D034281:Dynamins / Q000378:metabolism / Q000502:physiology*; D004705:Endocytosis / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D017434:Protein Structure, Tertiary / Q000235:genetics / Q000502:physiology; D011961:Receptors, Fc / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D020298:U937 Cells
D015703:Antigens, CD; D002966:Clathrin; C036832:Fc(alpha) receptor; D011961:Receptors, Fc; D034281:Dynamins