Maltseva Ekaterina A, Rechkunova Nadejda I, Sukhanova Maria V, Lavrik Olga I
From the Institute of Chemical Biology and Fundamental Medicine, 8 Lavrentiev Ave., 630090 Novosibirsk and.
From the Institute of Chemical Biology and Fundamental Medicine, 8 Lavrentiev Ave., 630090 Novosibirsk and the Department of Natural Sciences, Novosibirsk State University, 2 Pirogov St., 630090 Novosibirsk, Russia.
J Biol Chem. 2015 Sep 4;290(36):21811-20. doi: 10.1074/jbc.M115.646638. Epub 2015 Jul 13.
Poly(ADP-ribosyl)ation is a reversible post-translational modification that plays an essential role in many cellular processes, including regulation of DNA repair. Cellular DNA damage response by the synthesis of poly(ADP-ribose) (PAR) is mediated mainly by poly(ADP-ribose) polymerase 1 (PARP1). The XPC-RAD23B complex is one of the key factors of nucleotide excision repair participating in the primary DNA damage recognition. By using several biochemical approaches, we have analyzed the influence of PARP1 and PAR synthesis on the interaction of XPC-RAD23B with damaged DNA. Free PAR binds to XPC-RAD23B with an affinity that depends on the length of the poly(ADP-ribose) strand and competes with DNA for protein binding. Using (32)P-labeled NAD(+) and immunoblotting, we also demonstrate that both subunits of the XPC-RAD23B are poly(ADP-ribosyl)ated by PARP1. The efficiency of XPC-RAD23B PARylation depends on DNA structure and increases after UV irradiation of DNA. Therefore, our study clearly shows that XPC-RAD23B is a target of poly(ADP-ribosyl)ation catalyzed by PARP1, which can be regarded as a universal regulator of DNA repair processes.
聚(ADP-核糖)化是一种可逆的翻译后修饰,在许多细胞过程中发挥着重要作用,包括DNA修复的调控。通过合成聚(ADP-核糖)(PAR)介导的细胞DNA损伤反应主要由聚(ADP-核糖)聚合酶1(PARP1)介导。XPC-RAD23B复合物是参与初级DNA损伤识别的核苷酸切除修复的关键因素之一。通过使用几种生化方法,我们分析了PARP1和PAR合成对XPC-RAD23B与受损DNA相互作用的影响。游离的PAR以取决于聚(ADP-核糖)链长度的亲和力与XPC-RAD23B结合,并与DNA竞争蛋白质结合。使用(32)P标记的NAD(+)和免疫印迹,我们还证明XPC-RAD23B的两个亚基都被PARP1聚(ADP-核糖)化。XPC-RAD23B PARylation的效率取决于DNA结构,并在DNA紫外线照射后增加。因此,我们的研究清楚地表明,XPC-RAD23B是PARP1催化的聚(ADP-核糖)化的靶标,PARP1可被视为DNA修复过程的通用调节剂。