期刊
JOURNAL OF VIROLOGY
卷 83, 期 17, 页码 8893-8904出版社
AMER SOC MICROBIOLOGY
DOI: 10.1128/JVI.02239-08
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类别
资金
- National Institutes of Health [AI-13562]
- Ministry of Education, Science, Sports, Culture and Technology of Japan [20012056, 19041078, 20390137, 19590487]
- Research on Health Sciences focusing on Drug Innovation [SH54412]
- Ministry of Health, Labor and Welfare [19-01]
- Grants-in-Aid for Scientific Research [19041078, 19590487, 20012056] Funding Source: KAKEN
One of the two SP1 sites in the proximal enhancer of the human cytomegalovirus ( HCMV) major immediate-early (MIE) promoter is essential for transcription in human fibroblast cells (H. Isomura, M. F. Stinski, A. Kudoh, T. Daikoku, N. Shirata, and T. Tsurumi, J. Virol. 79: 9597-9607, 2005). Upstream of the two SP1 sites to -223 relative to the +1 transcription start site, there are an additional five DNA binding sites for eukaryotic transcription factors. We determined the effects of the various transcription factor DNA binding sites on viral MIE RNA transcription, viral gene expression, viral DNA synthesis, or infectious virus production. We prepared recombinant HCMV bacterial artificial chromosome (BAC) DNAs with either one site missing or one site present upstream of the two SP1 sites. Infectious recombinant HCMV BAC DNAs were transfected into various cell types to avoid the effect of the virion-associated transactivators. Regardless of the cell type, which included human fibroblast, endothelial, and epithelial cells, the CREB site had the most significant and independent effect on the MIE promoter. The other sites had a minor independent effect. However, the combination of the different transcription factor DNA binding sites was significantly stronger than multiple duplications of the CREB site. These findings indicate that the CREB site in the presence of the other sites has a major role for the replication of HCMV.
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