Sibbet G J, Cuthill S, Campo M S
Beatson Institute for Cancer Research, CRC Beatson Laboratories, Glasgow, Scotland.
J Virol. 1995 Jul;69(7):4006-11. doi: 10.1128/JVI.69.7.4006-4011.1995.
The minimal enhancer in the long control region of human papillomavirus type 16 regulates cell type and constitutive expression from the promoter P97. This region contains at least four DNase I footprints (fp4e, fp5e, fp6e, and fp7e). We have shown that fp5e is crucial to enhancer function and have described an apparently novel factor (PEF-1) binding fp5e (S. Cuthill, G. J. Sibbet, and M. S. Campo, Mol. Carcinog. 8:9-104, 1993). Further analyses reveal that Oct-1 or an Oct-related factor binds fp5e at a site overlapping that of PEF-1. The binding of Oct-1 to fp5e has been demonstrated by electrophoretic mobility shift assays, by oligonucleotide competition studies, and by using an Oct-1-specific anti-POU serum. The location of the Oct-1 site has been confirmed by a panel of mutants across fp5e. Mutations that block PEF-1 binding to fp5e also block enhancer/promoter activity of the long control region, whereas mutations that block Oct-1 binding significantly increase enhancer/promoter activity. Thus, although both PEF-1 and Oct-1 interact with fp5e, they appear to regulate human papillomavirus expression in opposite ways.
人乳头瘤病毒16型长控制区中的最小增强子调控细胞类型及启动子P97的组成型表达。该区域包含至少四个DNase I足迹(fp4e、fp5e、fp6e和fp7e)。我们已证明fp5e对增强子功能至关重要,并描述了一种明显新颖的因子(PEF-1)与fp5e结合(S. Cuthill、G. J. Sibbet和M. S. Campo,《分子致癌作用》8:9 - 104,1993年)。进一步分析表明,Oct-1或一个Oct相关因子在与PEF-1重叠的位点结合fp5e。通过电泳迁移率变动分析、寡核苷酸竞争研究以及使用Oct-1特异性抗POU血清,已证实Oct-1与fp5e的结合。通过一组跨越fp5e的突变体,已确认Oct-1位点的位置。阻断PEF-1与fp5e结合的突变也会阻断长控制区的增强子/启动子活性,而阻断Oct-1结合的突变则会显著增加增强子/启动子活性。因此,尽管PEF-1和Oct-1都与fp5e相互作用,但它们似乎以相反的方式调控人乳头瘤病毒的表达。