WRN helicase and FEN-1 form a complex upon replication arrest and together process branch-migrating DNA structures associated with the replication fork
WRN helicase and FEN-1 form a complex upon replication arrest and together process branch-migrating DNA structures associated with the replication fork
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DOI：
10.1091/mbc.e03-08-0567
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发表时间：
2004-02-01
影响因子：
3.3
通讯作者：
Brosh, RM
中科院分区：
文献类型：
--
作者：
Sharma, S;Otterlei, M;Brosh, RM
沃纳综合症是一种早衰性疾病，其特征是基因组不稳定、重组增加和复制缺陷。据推测，WRN 对某些复制叉结构的加工缺陷可能会导致基因组不稳定。荧光共振能量转移 (FRET) 分析表明，WRN 和 Flap Endonuclease-1 (FEN-1) 在体内形成复合物，共定位于与停滞的复制叉相关的病灶中。 WRN 有效刺激 FEN-1 对分支迁移双瓣结构的裂解，该结构是复制过程中 FEN-1 的生理底物。生化分析表明，WRN 解旋酶解旋与回归复制叉相关的鸡足 HJ 中间体，并刺激 FEN-1 以结构依赖性方式裂解解旋产物。这些结果为 WRN 和 FEN-1 之间的体内相互作用提供了证据，并表明这些蛋白质共同发挥作用来处理与复制叉相关的 DNA 结构。
Werner Syndrome is a premature aging disorder characterized by genomic instability, elevated recombination, and replication defects. It has been hypothesized that defective processing of certain replication fork structures by WRN may contribute to genomic instability. Fluorescence resonance energy transfer (FRET) analyses show that WRN and Flap Endonuclease-1 (FEN-1) form a complex in vivo that colocalizes in foci associated with arrested replication forks. WRN effectively stimulates FEN-1 cleavage of branch-migrating double-flap structures that are the physiological substrates of FEN-1 during replication. Biochemical analyses demonstrate that WRN helicase unwinds the chicken-foot HJ intermediate associated with a regressed replication fork and stimulates FEN-1 to cleave the unwound product in a structure-dependent manner. These results provide evidence for an interaction between WRN and FEN-1 in vivo and suggest that these proteins function together to process DNA structures associated with the replication fork.