ruva mutants that resolve holliday junctions but do not reverse replication forksruva突变体,解决霍利迪连接但不反向复制叉.pdfVIP

ruvA Mutants That Resolve Holliday Junctions but Do Not Reverse Replication Forks 1,2,3 4 1,2,3 4 ´ ´ Zeynep Baharoglu , Alison Sylvia Bradley , Marie Le Masson , Irina Tsaneva , Benedicte Michel1,2,3* ´ ´ ´ ´ ´ 1 CNRS, Centre de Genetique Moleculaire, UPR 2167, Gif-sur-Yvette, France, 2 Universite Paris-Sud, Orsay, France, 3 Universite Pierre et Marie Curie-Paris 6, Paris, France, 4 UCL Department of Biochemistry and Molecular Biology, University College London, London, United Kingdom Abstract RuvAB and RuvABC complexes catalyze branch migration and resolution of Holliday junctions (HJs) respectively. In addition to their action in the last steps of homologous recombination, they process HJs made by replication fork reversal, a reaction which occurs at inactivated replication forks by the annealing of blocked leading and lagging strand ends. RuvAB was recently proposed to bind replication forks and directly catalyze their conversion into HJs. We report here the isolation and characterization of two separation-of-function ruvA mutants that resolve HJs, based on their capacity to promote conjugational recombination and recombinational repair of UV and mitomycin C lesions, but have lost the capacity to reverse forks. In vivo and in vitro evidence indicate that the ruvA mutations affect DNA binding and the stimulation of RuvB helicase activity. This work shows that RuvA’s actions at forks and at HJs can be genetically separated, and that RuvA mutants compromised for fork reversal remain fully capable of homologous recombination.