title: A Luciferase Functional Quantitative Assay for Measuring NF-ĸB Promoter Transactivation Mediated by HTLV-1 and HTLV-2 Tax Proteins.
abstract: HTLV-1 and HTLV-2 viruses express Tax transactivator proteins required for viral genome transcription and capable of transforming cells in vivo and in vitro. Although Tax oncogenic potential needs to be further elucidated, it is well established that Tax proteins activate, among others, transcription factors of the NF-ĸB family, which are involved in immune and inflammatory responses, cell growth, apoptosis, stress responses and oncogenesis. Here, we describe a reporter gene assay applied for quantitative analysis of Tax-dependent NF-ĸB activation. The procedure is based on co-transfection of two individual vectors containing the cDNA for firefly and Renilla luciferase enzymes and vectors expressing Tax proteins. The luciferase expression is driven by cis-NF-ĸB promoter regulatory elements responsive to Tax transactivating factor. This assay is particularly useful to investigate Tax influence on NF-ĸB activation mediated by viral or host factors.
authors.affiliation: Section of Biology and Genetics, Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, Strata le Grazie 8, 37134, Verona, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Bergamo
authors.affiliation: Section of Biology and Genetics, Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, Strata le Grazie 8, 37134, Verona, Italy.
authors.forename: Erica
authors.initials: E
authors.lastname: Diani
authors.affiliation: Section of Biology and Genetics, Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, Strata le Grazie 8, 37134, Verona, Italy.
authors.forename: Umberto
authors.initials: U
authors.lastname: Bertazzoni
authors.affiliation: Section of Biology and Genetics, Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, Strata le Grazie 8, 37134, Verona, Italy. mariagrazia.romanelli@univr.it.
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Romanelli
mesh_terms: D016356:Gene Products, tax / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D015368:Human T-lymphotropic virus 1 / Q000235:genetics / Q000378:metabolism; D015367:Human T-lymphotropic virus 2 / Q000235:genetics / Q000378:metabolism; D006801:Humans; D049410:Luciferases, Renilla / Q000096:biosynthesis / Q000235:genetics; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D020218:Response Elements; D015533:Transcriptional Activation
chemical_list: D016356:Gene Products, tax; D016328:NF-kappa B; C514970:tax protein, Human T-lymphotrophic virus 1; C514971:tax protein, Human T-lymphotrophic virus 2; D049410:Luciferases, Renilla
keywords: Expressing vectors; Gene reporter assay; HTLV; Luciferase; NF-ĸB; Tax; Transfection