A bovine papillomavirus E1-related protein binds specifically to bovine papillomavirus DNA
A bovine papillomavirus E1-related protein binds specifically to bovine papillomavirus DNA
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牛乳头瘤病毒 E1 相关蛋白与牛乳头瘤病毒 DNA 特异性结合
作者：
Van G. Wilson;John H. Ludes
牛乳头瘤病毒（BPV）E1开放阅读框架基因在大肠杆菌中表达为RecA-E1融合蛋白。细菌表达的RecA-E1蛋白表现出序列特异性DNA结合活性;观察到与BPV基因组上核苷酸7819至93的区域（指定区域A）的强结合和与相邻的核苷酸7457至7818的区域（区域B）的弱结合。BPV衍生的RecA-E1蛋白和A区之间的相互作用似乎对BPV DNA具有高度特异性，因为与异源乳头瘤病毒DNA没有检测到类似的结合。通过在独特的HpaI位点消化A区，消除了与A区的结合，这表明RecA-E1结合位点位于或靠近HpaI识别序列。结合区域B，但不是区域A时，观察到从ID 13细胞的核提取物被用作E1蛋白的来源。ID 13提取物对A区结合的缺乏可能反映了E1 DNA结合活性的负调节。
The E1 open reading frame of bovine papillomavirus (BPV) was expressed as a RecA-E1 fusion protein in Escherichia coli. The bacterially expressed RecA-E1 protein exhibited sequence-specific DNA binding activity; strong binding to the region from nucleotides 7819 to 93 on the BPV genome (designated region A) and weak binding to the adjacent region from nucleotides 7457 to 7818 (region B) were observed. The interaction between the BPV-derived RecA-E1 protein and region A appeared to be highly specific for BPV DNA, as no comparable binding was detected with heterologous papillomavirus DNAs. Binding to region A was eliminated by digestion of region A at the unique HpaI site, which suggests that the RecA-E1 binding site(s) was at or near the HpaI recognition sequence. Binding to region B but not region A was observed when nuclear extracts from ID13 cells were used as a source of E1 proteins. The absence of region A binding by ID13 extracts may reflect a negative regulation of E1 DNA binding activity.
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