Cell Reports ( IF 7.5 ) Pub Date : 2020-07-07 , DOI: 10.1016/j.celrep.2020.107850 Anna Socha 1 , Di Yang 1 , Alicja Bulsiewicz 1 , Kelvin Yaprianto 1 , Marian Kupculak 1 , Chih-Chao Liang 1 , Andreas Hadjicharalambous 1 , Ronghu Wu 2 , Steven P Gygi 2 , Martin A Cohn 1
The Fanconi anemia (FA) pathway repairs DNA interstrand crosslinks (ICLs). Many FA proteins are recruited to ICLs in a timely fashion so that coordinated repair can occur. However, the mechanism of this process is poorly understood. Here, we report the purification of a FANCD2-containing protein complex with multiple subunits, including WRNIP1. Using live-cell imaging, we show that WRNIP1 is recruited to ICLs quickly after their appearance, promoting repair. The observed recruitment facilitates subsequent recruitment of the FANCD2/FANCI complex. Depletion of WRNIP1 sensitizes cells to ICL-forming drugs. We find that ubiquitination of WRNIP1 and the activity of its UBZ domain are required to facilitate recruitment of FANCD2/FANCI and promote repair. Altogether, we describe a mechanism by which WRNIP1 is recruited rapidly to ICLs, resulting in chromatin loading of the FANCD2/FANCI complex in an unusual process entailing ubiquitination of WRNIP1 and the activity of its integral UBZ domain.
中文翻译:
WRNIP1被招募到DNA链间交联并促进修复。
范可尼贫血(FA)途径可修复DNA链间交联(ICL)。及时将许多FA蛋白募集到ICL中,以便进行协调的修复。但是,对该过程的机制了解甚少。在这里,我们报告包含多个WWIP1多个亚基的含FANCD2的蛋白质复合物的纯化。使用活细胞成像,我们显示WRNIP1在出现后迅速被募集到ICL,从而促进修复。观察到的募集有助于随后募集FANCD2 / FANCI复合体。WRNIP1的耗尽会使细胞对ICL形成药物敏感。我们发现WRNIP1的泛素化及其UBZ域的活性是促进FANCD2 / FANCI募集和促进修复所必需的。总而言之,我们描述了一种将WRNIP1快速招募到ICL的机制,