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Originally published In Press as doi:10.1074/jbc.M108026200 on January 30, 2002

J. Biol. Chem., Vol. 277, Issue 14, 12023-12031, April 5, 2002
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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, and Shou-Jiang GaoDagger

From the Departments of Pediatrics and Microbiology, University of Texas Health Science Center at San Antonio, San Antonio, Texas 78229-3900

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.

* This work was supported in part by the Howard Hughes Medical Institute through the University of Texas Health Science Center at San Antonio for the Research Resource Program for Medical Schools S/G 0#7, the Elsa U. Pardee Foundation, Association for International Cancer Research, and National Institutes of Health Grant HL60604-01 (to S.-J. G.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Dept. of Pediatrics, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Dr., San Antonio, TX 78229-3900. Tel.: 210-567-5248; Fax: 210-567-6305; E-mail: gaos@uthscsa.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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