INDUCTION OF STRUCTURAL-CHANGES IN THE BOVINE PAPILLOMAVIRUS TYPE-1 ORIGIN OF REPLICATION BY THE VIRAL E1 AND E2 PROTEINS
INDUCTION OF STRUCTURAL-CHANGES IN THE BOVINE PAPILLOMAVIRUS TYPE-1 ORIGIN OF REPLICATION BY THE VIRAL E1 AND E2 PROTEINS
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DOI：
10.1073/pnas.91.19.8846
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发表时间：
1994-09-13
影响因子：
11.1
通讯作者：
BOROWIEC, JA
中科院分区：
文献类型：
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作者：
GILLETTE, TG;LUSKY, M;BOROWIEC, JA
化学和酶探针技术被用来研究牛乳头瘤病毒1型E1和E2蛋白与病毒复制起点（ori）的相互作用。E1被发现产生显着的扭曲的结构的ori，由高锰酸钾氧化的DNA测定。ori畸变的主要位点位于核心复制子序列的AT元件内和附近，尽管也检测到一些微小的结构转变。这些结构变化的诱导需要ATP，并且似乎需要ATP水解。E2被认为是减少的量，E1所需的ori失真，但并没有显着改变的模式的结构失真。与此相反，E2的存在下，导致在一个双相的机制E1结合到ori，核酸酶保护测定。在这些条件下，E1优先绑定到二分体对称区域包含保守的Hpa I网站。需要更高水平的E1结合到相邻的ori AT丰富的区域。因此，这些数据表明，E2可以命令E1的逐步结合到ori。
Chemical and enzymatic probing techniques were used to examine the interaction of the bovine papillomavirus type 1 E1 and E2 proteins with the viral origin of replication (ori). E1 was found to generate significant distortions to the structure of ori, as assayed by KMnO4 oxidation of DNA. The primary site of ori distortion was located within and adjacent to the AT-element of the core replicator sequence, although a number of minor structural transitions were also detected. The induction of these structural changes required ATP and appeared to require ATP hydrolysis. E2 was found to decrease the amount of E1 required for ori distortion but did not significantly alter the pattern of structural distortion. In contrast, the presence of E2 resulted in a biphasic mechanism for E1 binding to ori, as assayed by nuclease protection. Under these conditions, E1 bound preferentially to the dyad symmetry region containing the conserved Hpa I site. Higher levels of E1 were required for binding to the adjacent ori AT-rich region. Thus, these data suggest that E2 can order the stepwise binding of E1 to ori.