Tumor necrosis factor alpha induces the adenovirus early 3 promoter by activation of NF-kappaB.
The early transcription unit 3 (E3) of human adenoviruses encodes proteins which appear to subvert host defense mechanisms. For example, the E3/19K protein inhibits the transport of major histocompatibility complex (MHC) class I molecules to the cell surface and thereby prevents cell lysis by cytotoxic T cells. Tumor necrosis factor alpha (TNF) stimulates expression of MHC molecules on the cell surface of normal cells but not of E3(+) cells, rather, a further reduction of MHC expression is evident. This was attributed to the increased expression of E3/19K upon TNF treatment, an effect also observed for other E3 proteins. We investigated the mechanism of the TNF-mediated up-regulation of E3 products. We show that TNF stimulates expression of a luciferase reporter gene driven by the E3 promoter. Mutation of individual transcription factor binding sites within the E3 promoter reveals the importance of the NF-kappaB binding site kappa2 for TNF inducibility. Electrophoretic mobility shift assays using antibodies directed against various members of the NF-kappaB family demonstrate that stimulation by TNF is mediated by the p50-p65 NF-kappaB complex. TNF inducibility does not depend on coexpression of E1A and can be observed during infection. Interestingly, the E3 promoter seems to be the only early promoter responsive to TNF and the only adenovirus promoter containing an NF-kappaB site. The implications of this regulatory mechanism for the adenovirus life cycle and its pathogenesis are discussed.
Deryckere
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Hans-Spemann-Laboratorium, Max-Planck-Institut für Immunbiologie, Stübeweg 51, D-79108 Freiburg, Germany.
Burgert
H G
HG
D000256:Adenoviridae / Q000235:genetics*; D017500:Adenovirus E1A Proteins / Q000235:genetics; D017502:Adenovirus E2 Proteins / Q000235:genetics; D002460:Cell Line; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D016297:Mutagenesis, Site-Directed; D016328:NF-kappa B / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D015854:Up-Regulation
D017500:Adenovirus E1A Proteins; D017502:Adenovirus E2 Proteins; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha