Two different IFN-gamma nonresponsive variants derived from the B-cell lymphoma 70Z/3.
The kappa immunoglobulin (Igk) light chain locus is transcriptionally silent in the mouse B-cell lymphoma 70Z/3. However, exposure to lipopolysaccharide (LPS) or interferon-gamma (IFN) causes a marked increase in Igk transcription. By immunoselection, we isolated two variants that are nonresponsive to IFN. One variant, AT7.2, has retained its response to LPS (IFN-LPS+), whereas the other, AT3.3, is also nonresponsive to LPS (IFN-LPS-). Stable transfection of an intact Igk gene does not rescue the phenotype of either variant. Both variants have intact Igk genes and neither is deficient in the binding or uptake of IFN. Nuclear extracts from LPS-treated wild-type 70Z/3 cells show strong increases in three transcription factors: OTF-2, NF-kappa B, and kBF-A. Remarkably, when the IFN-LPS- variant is treated with LPS, all three transcription factors are still observed in the nuclear extracts. Treatment of wild-type cells with either LPS or IFN also causes a decrease in nuclear complexes that bind to two other regions of the Igk intron enhancer, the octenh and the E kappa MHCIC regions. Both of these changes are also observed after LPS or IFN treatment of the IFN-LPS- variant. Thus, this variant transduces the IFN and LPS signals at least into the nuclear compartment, but still fails to activate Igk transcription. In contrast, the IFN-LPS+ variant decreases neither the octenh nor the E kappa MHCIC binding complexes in response to IFN. This variant may be defective in transducing the IFN signal to the nucleus. These variants will be useful in studying the activation of Igk transcription and the IFN signaling pathway in B cells.
Department of Biological Structure, University of Washington, Seattle 98195.
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Rhodes
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Paull
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Sibley
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D001483:Base Sequence; D004247:DNA; D004268:DNA-Binding Proteins; D015536:Down-Regulation; D004742:Enhancer Elements, Genetic; D014644:Genetic Variation; D007145:Immunoglobulin kappa-Chains / Q000096:biosynthesis / Q000235:genetics; D007371:Interferon-gamma / Q000276:immunology* / Q000378:metabolism; D007438:Introns; D008070:Lipopolysaccharides / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism; D050813:Octamer Transcription Factor-2; D011485:Protein Binding; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; D007145:Immunoglobulin kappa-Chains; D008070:Lipopolysaccharides; D016328:NF-kappa B; D050813:Octamer Transcription Factor-2; C489206:Pou2f2 protein, mouse; D014157:Transcription Factors; D007371:Interferon-gamma; D004247:DNA; D013755:Tetradecanoylphorbol Acetate