PAPILLOMAVIRUS TRANSCRIPTIONAL PROGRAM

乳头瘤病毒DNA复制起点和E6/E7启动子 共享重叠序列。 此外，转录调控 蛋白质被证明是复制和复制所必需的 蛋白质调节转录。 这些监管区域已被 在两种病毒中分析：人乳头瘤病毒16型（HPV-16）和 牛乳头瘤病毒（BPV-1）。 HPV-16长控制区（LCR） 含有三个E2转录调节的结合位点， 地区 这些位点的突变分析表明， E2结合位点的数量与细胞增殖之间存在直接相关性。 SCC-9鳞状细胞癌中瞬时测定的DNA复制量 细胞癌细胞系在这些试验中，病毒E1和E2蛋白 从不同的表达载体表达。 P97突变 (E6/E7启动子）TATA盒对复制的影响最小，表明 这一区域的转录不是复制所必需的。 BPV-1的相应区域含有几个E2结合位点， 以及细胞转录因子包括SP1的结合位点， E2F、AP-1、YY-1和NF-1。 突变发生在这些位点， 在P89（E6/E7）启动子报告质粒的背景下。 的影响 通过瞬时转染牛胚胎检测这些突变 在存在或不存在E2蛋白的情况下，将E2蛋白与成纤维细胞（BEF）共培养。 我们 我以前已经表明，三个SP1网站需要两个基础 和反式激活表达。 对于NF-1观察到类似的结果 绝佳的价钱 YY-1和AP-1位点似乎也对 P89转录的调控。 然而，我们无法 证明了E2 F位点在我们的测定中的作用。

The papillomavirus origins of DNA replication and the E6/E7 promoters share overlapping sequences. In addition, transcriptional regulatory proteins have been shown to be required for replication and replication proteins modulate transcription. These regulatory regions have been analyzed in two viruses: human papillomavirus type 16 (HPV-16) and bovine papillomavirus (BPV-1). The HPV-16 long control region (LCR) contains three binding sites for the E2 transcriptional regulatory region. Mutational analysis of these sites demonstrated that there is a direct correlation between the number of E2 binding sites and the amount of DNA replication in transient assays in the SCC-9 squamous cell carcinoma cell line. In these assays the viral E1 and E2 proteins are expressed from separate expression vectors. Mutation of the P97 (E6/E7 promoter) TATA box had minimal effect on replication, indicating that transcription from this region is not required for replication. The corresponding region of BPV-1 contains several E2 binding sites as well as binding sites for cellular transcription factors including SP1, E2F, AP-1, YY-1, and NF-1. Mutations were made in these sites in the context of a P89 (E6/E7) promoter reporter plasmid. The effects of these mutations were assayed by transient transfection in bovine embryo fibroblasts (BEF) in the presence or absence of the E2 protein. We have previously shown that three SP1 sites are required for both basal and transactivated expression. Similar results were seen for the NF-1 site. The YY-1 and AP-1 sites also appear to be important for regulation of P89 transcription. However, we have been unable to demonstrate a role for the E2F site in our assays.