Eggleston A K, Mitchell A H, West S C
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, United Kingdom.
Cell. 1997 May 16;89(4):607-17. doi: 10.1016/s0092-8674(00)80242-1.
Purified proteins have been used to reconstitute an in vitro system for the medial-to-late stages of recombination in E. coli. In this system, RecA protein formed recombination intermediates that were processed by the actions of the RuvA, RuvB, and RuvC proteins. RuvAB was found to promote branch migration, to dissociate the RecA filament, and to modulate the orientation of cleavage of Holliday junction resolution by RuvC. Monoclonal antibodies directed against RuvA, RuvB, or RuvC inhibited resolution in the reconstituted system. Specific protein-protein interactions between the branch migration motor (RuvB) and the resolvase (RuvC) were also observed. These results provide evidence for coordinated action during the late stages of recombination, possibly involving the assembly of a RuvABC branch migration/resolution complex.
纯化的蛋白质已被用于重建一个用于大肠杆菌中重组中期到后期阶段的体外系统。在这个系统中，RecA蛋白形成了重组中间体，这些中间体通过RuvA、RuvB和RuvC蛋白的作用进行加工。发现RuvAB促进分支迁移、解离RecA丝，并调节RuvC对霍利迪连接点切割的方向。针对RuvA、RuvB或RuvC的单克隆抗体在重建系统中抑制切割。还观察到分支迁移马达（RuvB）和解旋酶（RuvC）之间存在特定的蛋白质-蛋白质相互作用。这些结果为重组后期的协同作用提供了证据，可能涉及RuvABC分支迁移/切割复合物的组装。
