Parsons C A, Tsaneva I, Lloyd R G, West S C
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Herts, United Kingdom.
Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5452-6. doi: 10.1073/pnas.89.12.5452.
The RuvA, RuvB, and RuvC proteins of Escherichia coli are required for the recombinational repair of ultraviolet light- or chemical-induced DNA damage. In vitro, RuvC protein interacts with Holliday junctions in DNA and promotes their resolution by endonucleolytic cleavage. In this paper, we investigate the interaction of RuvA and RuvB proteins with model Holliday junctions. Using band-shift assays, we show that RuvA binds synthetic Holliday structures to form specific protein-DNA complexes. Moreover, in the presence of ATP, the RuvA and RuvB proteins act in concert to promote dissociation of the synthetic Holliday structures. The dissociation reaction requires both RuvA and RuvB and a nucleotide cofactor (ATP or dATP) and is rapid (40% of DNA molecules dissociate within 1 min). The reaction does not occur when ATP is replaced by either ADP or the nonhydrolyzable analog of ATP, adenosine 5'-[gamma-thio]triphosphate. We suggest that the RuvA and RuvB proteins play a specific role in the branch migration of Holliday junctions during postreplication repair of DNA damage in E. coli.
大肠杆菌的RuvA、RuvB和RuvC蛋白是紫外线或化学物质诱导的DNA损伤进行重组修复所必需的。在体外，RuvC蛋白与DNA中的霍利迪连接体相互作用，并通过核酸内切酶切割促进其解离。在本文中，我们研究了RuvA和RuvB蛋白与模型霍利迪连接体的相互作用。使用凝胶迁移试验，我们发现RuvA结合合成的霍利迪结构以形成特定的蛋白质-DNA复合物。此外，在ATP存在的情况下，RuvA和RuvB蛋白协同作用以促进合成霍利迪结构的解离。解离反应需要RuvA和RuvB以及核苷酸辅因子（ATP或dATP），并且速度很快（40%的DNA分子在1分钟内解离）。当ATP被ADP或ATP的不可水解类似物腺苷5'-[γ-硫代]三磷酸取代时，反应不会发生。我们认为，RuvA和RuvB蛋白在大肠杆菌DNA损伤复制后修复过程中霍利迪连接体的分支迁移中发挥特定作用。
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