Induction of the human protein P56 by interferon, double-stranded RNA, or virus infection.
P56 is the most abundant protein induced by interferon (IFN) treatment of human cells. To facilitate studies on its induction pattern and cellular functions, we expressed recombinant P56 as a hexahistidine-tagged protein in Escherichia coli and purified it to apparent homogeneity using affinity chromatography. A polyclonal antibody raised against this recombinant protein was used to show that P56 is primarily a cytoplasmic protein. Cellular expression of P56 by transfection did not inhibit the replication of vesicular stomatitis virus and encephalomyocarditis virus. P56 synthesis was rapidly induced by IFN-beta, and the protein had a half-life of 6 h. IFN-gamma or poly(A)(+) could not induce the protein, but poly(I)-poly(C) or an 85-bp synthetic double-stranded RNA efficiently induced it. Similarly, infection of GRE cells, which are devoid of type I IFN genes, by vesicular stomatitis virus, encephalomyocarditis virus, or Sendai virus caused P56 induction. Surprisingly, Sendai virus could also induce P56 in the mutant cell line P2.1, which cannot respond to either IFN-alpha/beta or double-stranded RNA. Induction of P56 in the P2.1 cells and the parental U4C cells by virus infection was preceded by activation of IRF-3 as judged by its translocation to the nucleus from the cytoplasm.
Guo
J
J
Department of Molecular Biology, The Lerner Research Institute, The Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, Ohio 44195, USA.
Peters
K L
KL
Sen
G C
GC
D000911:Antibodies, Monoclonal / Q000276:immunology; D002460:Cell Line; D004686:Encephalomyocarditis virus / Q000254:growth & development; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D016899:Interferon-beta / Q000494:pharmacology; D007372:Interferons / Q000494:pharmacology*; D012330:RNA, Double-Stranded / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000235:genetics; D018108:Respirovirus / Q000254:growth & development; D014157:Transcription Factors / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D014721:Vesicular stomatitis Indiana virus / Q000254:growth & development
D000911:Antibodies, Monoclonal; D012330:RNA, Double-Stranded; D011994:Recombinant Proteins; D014157:Transcription Factors; D016899:Interferon-beta; D007372:Interferons