Bream G L, Vaillancourt P, Botchan M R
Department of Molecular and Cell Biology, University of California, Berkeley 94720.
J Virol. 1992 Dec;66(12):7319-27. doi: 10.1128/JVI.66.12.7319-7327.1992.
The bovine papillomavirus upstream regulatory region represents a common element in the regulation of transcription from the five early viral promoters. We have determined the sequences required for transcription from the viral P1 promoter, which is located at the 5' end of the upstream regulatory region. In vitro transcription from P1 requires a 123-bp fragment (nucleotides 7153 to 7275; -33 to +90) consisting of an upstream TATA-like sequence as well as an unidentified protein which binds to sequences immediately downstream of the initiation site. In vivo, this promoter requires additional downstream sequences (to position +160; nucleotide 7345) for maximal activity but does not require any additional DNA sequence upstream of a putative TATA box. Four regions within the downstream sequence from +9 to +160 are protected from DNase I digestion by proteins present in a HeLa cell extract. The presence of these sites correlates with the level of P1 activity. A constitutive enhancer maps to this same region, and mutations in this enhancer have been shown to affect downstream promoters. Deletion analysis indicates that the same sequences are required by both the P1 promoter and the constitutive enhancer, suggesting that the same proteins function in both activities.
牛乳头瘤病毒上游调控区是调控五个早期病毒启动子转录的共同元件。我们已经确定了位于上游调控区5'端的病毒P1启动子转录所需的序列。P1的体外转录需要一个123 bp的片段(核苷酸7153至7275;-33至+90),该片段由一个上游类TATA序列以及一种与起始位点下游紧邻序列结合的未知蛋白质组成。在体内,该启动子需要额外的下游序列(至+160位;核苷酸7345)以实现最大活性,但在假定的TATA框上游不需要任何额外的DNA序列。从+9至+160的下游序列中的四个区域受到HeLa细胞提取物中存在的蛋白质对DNase I消化的保护。这些位点的存在与P1活性水平相关。一个组成型增强子定位于同一区域,并且已证明该增强子中的突变会影响下游启动子。缺失分析表明,P1启动子和组成型增强子都需要相同的序列,这表明相同的蛋白质在这两种活性中发挥作用。