Rawlins D R, Rosenfeld P J, Wides R J, Challberg M D, Kelly T J
Cell. 1984 May;37(1):309-19. doi: 10.1016/0092-8674(84)90327-1.
Efficient initiation of adenovirus DNA replication requires the presence of specific terminal nucleotide sequences that collectively constitute the viral origin of replication. Using plasmids with deletions or base substitutions in a cloned segment of DNA derived from the terminus of the adenovirus 2 genome, we have demonstrated that the origin contains two functionally distinct regions. The first 18 bp of the viral genome are sufficient to support a limited degree of initiation. However, the presence of a sequence in the region between nucleotides 19 and 67 greatly enhances the efficiency of the initiation reaction. This region contains a specific binding site for a protein present in uninfected cells (KD = 2 X 10(-11) M). The bound protein protects the DNA segment between base pairs 19 and 43 from attack by DNAase I. Studies with deletion mutants indicate that binding of the cellular protein is responsible for the enhancement of initiation.
腺病毒DNA复制的有效起始需要特定的末端核苷酸序列的存在，这些序列共同构成病毒复制起点。利用在源自腺病毒2基因组末端的一段克隆DNA片段中存在缺失或碱基替换的质粒，我们已经证明该起点包含两个功能不同的区域。病毒基因组的前18个碱基对足以支持有限程度的起始。然而，在核苷酸19至67之间区域中一个序列的存在极大地提高了起始反应的效率。该区域包含未感染细胞中存在的一种蛋白质的特异性结合位点（解离常数KD = 2×10⁻¹¹ M）。结合的蛋白质保护碱基对19至43之间的DNA片段免受DNA酶I的攻击。对缺失突变体的研究表明，细胞蛋白质的结合是起始增强的原因。
