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Journal of Virology, March 2000, p. 2459-2465, Vol. 74, No. 5
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Adeno-Associated Virus Major Rep78 Protein Disrupts Binding of TATA-Binding Protein to the p97 Promoter of Human Papillomavirus Type 16dagger

Pei-Fen Su,1,2 Shu-Yuan Chiang,1 Cheng-Wen Wu,1,* and Felicia Y.-H. Wu1

Division of Cancer Research, Institute of Biomedical Sciences, Academia Sinica,1 and Institute of Life Sciences, National Defense Medical Center,2 Taipei, Taiwan, Republic of China

Received 24 June 1999/Accepted 7 December 1999

Adeno-associated virus type 2 (AAV) is known to inhibit the promoter activities of several oncogenes and viral genes, including the human papillomavirus type 16 (HPV-16) E6 and E7 transforming genes. However, the target elements of AAV on the long control region (LCR) upstream of E6 and E7 oncogenes are elusive. A chloramphenicol acetyltransferase assay was performed to study the effect of AAV on the transcription activity of the HPV-16 LCR in SiHa (HPV-positive) and C-33A (HPV-negative) cells. The results reveal that (i) AAV inhibited HPV-16 LCR activity in a dose-dependent manner, (ii) AAV-mediated inhibition did not require the HPV gene products, and (iii) the AAV replication gene product Rep78 was involved in the inhibition. Deletion mutation analyses of the HPV-16 LCR showed that regulatory elements outside the core promoter region of the LCR may not be direct targets of AAV-mediated inhibition. Further study with the electrophoretic mobility shift assay demonstrated that Rep78 interfered with the binding of TATA-binding protein (TBP) to the TATA box of the p97 core promoter more significantly than it disrupted the preformed TBP-TATA complex. These data thus suggest that Rep78 may inhibit transcription initiation of the HPV-16 LCR by disrupting the interaction between TBP and the TATA box of the p97 core promoter.

* Corresponding author. Mailing address: Division of Cancer Research, Institute of Biomedical Sciences, Academia Sinica, Nankang, Taipei 115, Taiwan, Republic of China. Phone: 886-2-2653-4401, ext. 8002. Fax: 886-2-2782-9142. E-mail: Kenwu{at}nhri.org.tw.

dagger This paper is dedicated to the memory of Felicia Wu, who died 19 July 1999.

Journal of Virology, March 2000, p. 2459-2465, Vol. 74, No. 5
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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