Ligand-induced phosphorylation of the human interferon-gamma receptor. Dependence on the presence of a functionally active receptor.
Phosphorylation of the human interferon-gamma (IFN gamma) receptor was studied in three cell lines of distinct lineages using radiophosphate labeling techniques. Receptors from unstimulated Colo-205 displayed a low level of constitutive phosphorylation which was enhanced 5.3-fold following exposure of the cells to either purified recombinant human IFN gamma or phorbol myristate acetate. Enhanced receptor phosphorylation was specific, dose- and time-dependent, reversible, and affected only serine and threonine residues. Increased phosphorylation was observed only when cells were treated with human IFN gamma or phorbol myristate acetate and not with murine IFN gamma, human IFN alpha, human tumor necrosis factor-alpha, or epidermal growth factor. The biologic relevance of IFN gamma receptor phosphorylation was suggested by three additional observations; 1) there was a close correlation between the extent of receptor phosphorylation and the magnitude of the cellular response induced, 2) TNF alpha concomitantly enhanced both IFN gamma-dependent HLA-DR expression and IFN gamma-dependent receptor phosphorylation on Colo-205, and 3) phosphorylation of functionally inactive recombinant murine IFN gamma receptors expressed on transfected human 293 or Colo-205 cells was not induced by murine IFN gamma but was induced by the homologous human ligand. These results suggest that phosphorylation of the IFN gamma receptor is an important step in the development of IFN gamma-dependent cellular responses and indicates that phosphorylation requires a functionally active receptor.
Hershey
G K
GK
Department of Pathology, Washington University School of Medicine, St. Louis, Missouri.
McCourt
D W
DW
Schreiber
R D
RD
D000818:Animals; D002460:Cell Line; D006684:HLA-DR Antigens / Q000235:genetics; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D051379:Mice; D010766:Phosphorylation; D011971:Receptors, Immunologic / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D017471:Receptors, Interferon; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection
D006684:HLA-DR Antigens; D008024:Ligands; D011971:Receptors, Immunologic; D017471:Receptors, Interferon; D007371:Interferon-gamma; D013755:Tetradecanoylphorbol Acetate