Li R, Botchan M R
Department of Molecular and Cell Biology University of California, Berkeley 94720.
Cell. 1993 Jun 18;73(6):1207-21. doi: 10.1016/0092-8674(93)90649-b.
For papillomavirus DNA replication, the E2 enhancer protein cooperatively assists in binding of the E1 helicase to the origin. We report that, at limiting E1 and E2 levels, the enhancer proteins GAL4-VP16 and GAL4-p53(1-73) stimulate BPV in vitro DNA replication. This cell-free system was used to ascertain whether the acidic activation domains have a cellular target important for replication. Cellular extracts were depleted of replication activity by passage through a VP16 affinity column. The protein depleted was the cellular factor replication protein A. The direct interaction between replication protein A and VP16, as well as the activation of replication by VP16, is dependent upon the C-terminus of the VP16 activation domain. E2 and the activation domain of p53 also interact with replication protein A. We suggest that a link between transcription and replication involves factors that help convert a closed DNA complex to an open complex.
对于乳头瘤病毒DNA复制，E2增强子蛋白协同辅助E1解旋酶与起始位点的结合。我们报告称，在E1和E2水平有限的情况下，增强子蛋白GAL4-VP16和GAL4-p53(1-73)可刺激体外BPV DNA复制。该无细胞系统用于确定酸性激活结构域是否具有对复制很重要的细胞靶点。通过VP16亲和柱处理使细胞提取物的复制活性耗尽。耗尽的蛋白是细胞因子复制蛋白A。复制蛋白A与VP16之间的直接相互作用以及VP16对复制的激活取决于VP16激活结构域的C末端。E2和p53的激活结构域也与复制蛋白A相互作用。我们认为转录与复制之间的联系涉及有助于将闭合DNA复合物转化为开放复合物的因子。
