tailieunhanh - Báo cáo khoa học: Transcription factor human Skn-1a enhances replication of human papillomavirus DNA through the direct binding to two sites near the viral replication origin

Human papillomavirus type 16 (HPV16) DNA replication, which requires two viral proteins E1 and E2, occurs only in the differentiating epithelium. Besides the general factors necessary for cellular DNA synthesis, other unidentified cellular factors are assumed to be involved in the regulation of HPV DNA replication. | ỊFEBS Journal Transcription factor human Skn-1a enhances replication of human papillomavirus DNA through the direct binding to two sites near the viral replication origin Iwao Kukimoto Seiichiro Mori Hidetaka Sato Takamasa Takeuchi and Tadahito Kanda Center for Pathogen Genomics National institute of Infectious Diseases Tokyo Japan Keywords DNA replication hSkn-1a human papillomavirus keratinocyte differentiation transcription factor Correspondence I. Kukimoto Center for Pathogen Genomics National Institute of Infectious Diseases 1-23-1 Toyama Shinjuku-ku Tokyo 162-8640 Japan Fax 81 3 5285 1166 Tel 81 3 5285 1111 E-mail ikuki@ Received 10 December 2007 revised 19 March 2008 accepted 15 April 2008 doi Human papillomavirus type 16 HPV16 DNA replication which requires two viral proteins E1 and E2 occurs only in the differentiating epithelium. Besides the general factors necessary for cellular DNA synthesis other unidentified cellular factors are assumed to be involved in the regulation of HPV DNA replication. In the present study we found that the POU-domain transcription factor human Skn-1a which induces the terminal differentiation of keratinocytes and activates the HPV16 late promoter enhanced the transient replication of a plasmid containing the HPV16 replication origin in HEK293 cells when co-transfected with a plasmid expressing E1 and E2. An electrophoretic mobility shift assay with a bacte-rially expressed human Skn-1a or an extract of HeLa cells over-expressing human Skn-1a revealed the presence of two human Skn-1a binding sites that are distinct from the known three sites near the replication origin. A chromatin immunoprecipitation analysis showed that human Skn-1a bound to these sites in cells. Nucleotide substitutions in the sites abolished the binding of human Skn-1a and the human Skn-1a-mediated replication enhancement. The data strongly suggest that through the binding to the two sites human Skn-1a enhances HPV

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