Production of a monoclonal antibody directed against an interferon-induced 56,000-dalton protein and its use in the study of this protein.
Interferon (IFN) treatment of cells induces the synthesis of several new proteins. A hybridoma cell line producing monoclonal antibody to the IFN-induced 56,000-dalton protein has been developed. The IFN-induced 56,000-dalton protein is synthesized by a variety of different cells and in response to IFN-alpha, IFN-beta, and IFN-gamma. The induction of this protein is dependent on de novo RNA synthesis, since its induction is inhibited if actinomycin D and the IFNs are added to the cells simultaneously. Labeling of IFN-treated cells at 4-h intervals at various times after the addition of the IFNs reveals that the synthesis of the 56,000-dalton protein in IFN-alpha-treated cells peaks within 12 h after the addition of the IFN and is no longer enhanced 20 h after exposure to the IFN. In contrast, IFN-gamma-treated cells continue to show an enhanced synthesis of this IFN-induced protein even after 20 h of exposure to the IFN. Thus, the synthesis of the IFN-induced 56,000-dalton protein is regulated differently by the different IFNs. When cells are treated with IFN-alpha or IFN-gamma in the presence of cycloheximide, and actinomycin D is added prior to the removal of the cycloheximide, the cells produce the IFN-induced 56,000-dalton protein and develop an antiviral state in response to both IFN-alpha and IFN-gamma. These results demonstrate that the synthesis of the 56,000-dalton protein is not dependent on the synthesis of an intermediary protein and that the establishment of an antiviral state occurs in the absence of multiple transcriptional events.
D048868:Adaptor Proteins, Signal Transducing; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002460:Cell Line; D011232:Chemical Precipitation; D003609:Dactinomycin / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007163:Immunosorbent Techniques; D066298:In Vitro Techniques; D007372:Interferons / Q000494:pharmacology*; D007526:Isoelectric Point; D008970:Molecular Weight; D011506:Proteins / Q000235:genetics / Q000276:immunology*; D016601:RNA-Binding Proteins; D013997:Time Factors; D014762:Viral Interference / Q000187:drug effects
D048868:Adaptor Proteins, Signal Transducing; D000911:Antibodies, Monoclonal; C484290:IFIT1 protein, human; D011506:Proteins; D016601:RNA-Binding Proteins; D003609:Dactinomycin; D007372:Interferons