Transcriptional regulation of Kaposi's sarcoma-associated herpesvirus-encoded oncogene viral interferon regulatory factor by a novel transcriptional silencer, Tis

Xin Ping Wang, Yan Jin Zhang, Jian Hong Deng, Hong Yi Pan, Fu Chun Zhou, Shou Jiang Gao

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Viral interferon regulatory factor (vIRF) encoded by Kaposi's sarcoma-associated herpesvirus (KSHV) has been shown to transform NIH3T3 and Rat-1 cells, inhibit interferon signal transduction, and regulate the expression of KSHV genes. We had previously characterized the vIRF core promoter and defined a 12-O-tetradecanoylphorbol-13-acetate (TPA)-responsive region in the upstream regulatory sequence of vIRF gene. Here, we have further identified a novel transcriptional silencer, named Tis in this region. Tis represses the promoter activities of vIRF and heterologous herpes simplex virus thymidine kinase genes in both position- and orientation-independent manners. Deletion analysis has identified a cis-element of 23 nucleotides that is essential for the negative regulation. Two Tis-binding protein complexes, named vR1 and vR2, were observed by electrophoretic mobility shift assays using nuclear extracts from both KSHV-negative and -positive cell lines. A sequence fragment GAGTTAATAGGTAGAG in the cis-element was shown to be required for the DNA-protein interactions as well as the repression of vIRF promoter activity. Point-mutation analysis identified TTAAT and GTTAATAG as the core sequence motifs for the binding of vR1 and vR2, respectively. These results define the function of a novel transcriptional silencer in the regulation of vIRF gene expression.

Original languageEnglish (US)
Pages (from-to)12023-12031
Number of pages9
JournalJournal of Biological Chemistry
Volume277
Issue number14
DOIs
StatePublished - Apr 5 2002

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Human Herpesvirus 8
Oncogenes
Genes
Regulator Genes
Electrophoretic mobility
Signal transduction
Thymidine Kinase
Electrophoretic Mobility Shift Assay
Tetradecanoylphorbol Acetate
Simplexvirus
Viruses
Point Mutation
Gene expression
Interferons
viral interferon regulatory factors
Rats
Assays
Signal Transduction
Carrier Proteins
Acetates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Transcriptional regulation of Kaposi's sarcoma-associated herpesvirus-encoded oncogene viral interferon regulatory factor by a novel transcriptional silencer, Tis. / Wang, Xin Ping; Zhang, Yan Jin; Deng, Jian Hong; Pan, Hong Yi; Zhou, Fu Chun; Gao, Shou Jiang.

In: Journal of Biological Chemistry, Vol. 277, No. 14, 05.04.2002, p. 12023-12031.

Research output: Contribution to journalArticle

Wang, Xin Ping ; Zhang, Yan Jin ; Deng, Jian Hong ; Pan, Hong Yi ; Zhou, Fu Chun ; Gao, Shou Jiang. / Transcriptional regulation of Kaposi's sarcoma-associated herpesvirus-encoded oncogene viral interferon regulatory factor by a novel transcriptional silencer, Tis. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 14. pp. 12023-12031.
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abstract = "Viral interferon regulatory factor (vIRF) encoded by Kaposi's sarcoma-associated herpesvirus (KSHV) has been shown to transform NIH3T3 and Rat-1 cells, inhibit interferon signal transduction, and regulate the expression of KSHV genes. We had previously characterized the vIRF core promoter and defined a 12-O-tetradecanoylphorbol-13-acetate (TPA)-responsive region in the upstream regulatory sequence of vIRF gene. Here, we have further identified a novel transcriptional silencer, named Tis in this region. Tis represses the promoter activities of vIRF and heterologous herpes simplex virus thymidine kinase genes in both position- and orientation-independent manners. Deletion analysis has identified a cis-element of 23 nucleotides that is essential for the negative regulation. Two Tis-binding protein complexes, named vR1 and vR2, were observed by electrophoretic mobility shift assays using nuclear extracts from both KSHV-negative and -positive cell lines. A sequence fragment GAGTTAATAGGTAGAG in the cis-element was shown to be required for the DNA-protein interactions as well as the repression of vIRF promoter activity. Point-mutation analysis identified TTAAT and GTTAATAG as the core sequence motifs for the binding of vR1 and vR2, respectively. These results define the function of a novel transcriptional silencer in the regulation of vIRF gene expression.",
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