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J Interferon Cytokine Res ; 21(11): 981-90, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11747630

ABSTRACT

To elucidate the host cell defense mechanisms in response to Sindbis viral infection, we have started to characterize interferon (IFN)-stimulated response element (ISRE)-binding proteins activated in infected cells that are involved in the transcriptional induction of IFN type I-inducible genes. Using electromobility shift assays (EMSA), we detected several protein complexes with a human IFN-stimulated gene 15 (ISG15) ISRE in extracts from virus-infected L929 cells that were absent in extracts from uninfected cells. Comigration with Newcastle disease virus-activated ISRE-binding complexes, ISRE-binding specificity, supershift experiments, and conditions of formation indicate that the complexes activated by Sindbis viral infection in L929 cells correspond to DRAF1 and ISG factor 3 (ISGF3). Transfection of L929 cells with poly rI:rC induced only ISGF3. DRAF1 could be detected in Sindbis virus-infected mouse embryo fibroblasts derived from IFNR type I and type II KO mice. Viral RNA synthesis is required for activation of DRAF1.


Subject(s)
DNA-Binding Proteins/metabolism , Interferon Type I/pharmacology , Response Elements , Sindbis Virus/physiology , Transcription Factors/metabolism , Animals , Cell Line , Chlorocebus aethiops , Cytokines/biosynthesis , Cytokines/genetics , Dactinomycin/pharmacology , Electrophoretic Mobility Shift Assay , Macromolecular Substances , Mice , Nucleic Acid Synthesis Inhibitors/pharmacology , RNA, Double-Stranded/pharmacology , RNA, Messenger/biosynthesis , RNA, Viral/biosynthesis , Sindbis Virus/genetics , Transcription Factors/genetics , Transcriptional Activation , Ubiquitins , Vero Cells
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