Effects of subunit mutation on the localization to coated pits and internalization of cross-linked IgE-receptor complexes.
IgE receptors of mast cells, Fc epsilon RI, localize to coated pits and internalize after cross-linking. We investigated whether any one of the receptor's four distinctive cytoplasmic domains regulates these phenomena. COS cells, which lack Fc epsilon RI entirely, and P815 mouse mastocytoma cells that lack the alpha and beta subunits of the tetrameric Fc epsilon RI (alpha beta gamma 2), were transfected with wild-type, incomplete, or variant Fc epsilon RI. IgE-receptor complexes were observed by electron microscopy. Before cross-linking with anti-IgE gold particles, receptors were not preferentially localized to coated pits, which occupy approximately 1% of the cell surface. After cross-linking, up to 10 to 20% of the wild-type and most other receptor variants were in coated pits in transfected P815 cells at any one time. beta-less variants localized normally but, surprisingly, receptors containing a variant beta subunit showed reduced localization. "Receptors" consisting simply of the lipid-anchored ectodomains of the human alpha subunit failed to localize to coated pits. In general, cross-linked receptors that localized to coated pits were progressively internalized, whereas receptors that failed to accumulate in coated pits were not. We conclude that no single cytoplasmic domain of the Fc epsilon RI uniquely controls its ligand-induced localization to coated pits and internalization.
Mao
S Y
SY
Section on Chemical Immunology, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD 20892.
Pfeiffer
J R
JR
Oliver
J M
JM
Metzger
H
H
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003034:Coated Pits, Cell-Membrane / Q000737:chemistry* / Q000648:ultrastructure; D004705:Endocytosis; D012515:Mast-Cell Sarcoma / Q000737:chemistry; D051379:Mice; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009154:Mutation; D011817:Rabbits; D051381:Rats; D017455:Receptors, IgE / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D017455:Receptors, IgE