Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA.
Curr Opin Chem Biol. 2011 Oct;15(5):580-6. doi: 10.1016/j.cbpa.2011.07.024. Epub 2011 Sep 9.
The replication system of phage T7 provides a model for DNA replication. Biochemical, structural, and single-molecule analyses together provide insight into replisome mechanics. A complex of polymerase, a processivity factor, and helicase mediates leading strand synthesis. Establishment of the complex requires an interaction of the C-terminal tail of the helicase with the polymerase. During synthesis the complex is stabilized by other interactions to provide for a processivity of 5 kilobase (kb). The C-terminal tail also interacts with a distinct region of the polymerase to captures dissociating polymerase to increase the processivity to >17kb. The lagging strand is synthesized discontinuously within a loop that forms and resolves during each cycle of Okazaki fragment synthesis. The synthesis of a primer as well as the termination of a fragment signal loop resolution.
噬菌体 T7 的复制系统为 DNA 复制提供了一个模型。生化、结构和单分子分析共同提供了对复制体机制的深入了解。聚合酶、持续因子和解旋酶的复合物介导了前导链的合成。复合物的建立需要解旋酶的 C 末端尾巴与聚合酶的相互作用。在合成过程中,复合物通过其他相互作用稳定,从而提供 5 千碱基(kb)的持续合成。C 末端尾巴还与聚合酶的一个独特区域相互作用,捕获解离的聚合酶以提高持续合成的效率至>17kb。滞后链在每个冈崎片段合成循环中形成和解决的环内不连续合成。引物的合成以及片段终止信号环的解决都依赖于该过程。