Abstract
The role of Vav in the transcriptional regulation of the human interferon-gamma (IFN-gamma) promoter was investigated. Overexpression of Vav in Jurkat-TAg cells enhanced T cell receptor (TCR)-induced activation of a luciferase (Luc) reporter gene construct driven by cis-regulatory element of the IFN-gamma gene (-346 to +7). Electrophoresis mobility shift and Luc reporter assays demonstrated that the DNA-binding and transcriptional activity of the proximal AP-1-dependent NFAT site (positions -172 to -138), the AP-1/Ying-Yang 1 (YY1)-binding site (-209 to -184), and a consensus AP-1-binding site were upregulated by Vav. Vav enhanced TCR-induced activation of c-Jun N-terminal kinase (JNK) and its upstream regulator, Rho family GTPases. Finally, coexpression of a dominant-negative Rac1 mutant suppressed Vav-mediated upregulation of the transcriptional and DNA-binding activity of the proximal NFAT/AP-1 site and the AP-1/YY1 site, as well as the complete IFN-gamma promoter activity. Vav activates the IFN-gamma promoter via upregulation of AP-1-binding through a Rac1/JNK pathway.
MeSH terms
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DNA-Binding Proteins / metabolism
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Dose-Response Relationship, Drug
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Enzyme Activation / drug effects
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Gene Expression / drug effects
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Genes, Reporter
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Humans
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Interferon-gamma / genetics*
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JNK Mitogen-Activated Protein Kinases
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Jurkat Cells / metabolism
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Mitogen-Activated Protein Kinases / metabolism
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NFATC Transcription Factors
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Nuclear Proteins*
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Oncogene Proteins / genetics
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Oncogene Proteins / metabolism*
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Oncogene Proteins / pharmacology
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Promoter Regions, Genetic / drug effects
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Promoter Regions, Genetic / physiology*
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Proto-Oncogene Proteins c-vav
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Receptors, Antigen, T-Cell / metabolism
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Response Elements / drug effects
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Response Elements / physiology
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Transcription Factor AP-1 / metabolism*
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Transcription Factors / metabolism
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Transfection
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Up-Regulation / drug effects
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Up-Regulation / physiology*
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rho GTP-Binding Proteins / metabolism
Substances
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DNA-Binding Proteins
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NFATC Transcription Factors
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Nuclear Proteins
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Oncogene Proteins
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Proto-Oncogene Proteins c-vav
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Receptors, Antigen, T-Cell
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Transcription Factor AP-1
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Transcription Factors
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VAV1 protein, human
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Interferon-gamma
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JNK Mitogen-Activated Protein Kinases
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Mitogen-Activated Protein Kinases
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rho GTP-Binding Proteins