Department of Human Molecular Genetics and Biochemistry, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel.
Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER) and Department of Genetics, University of Sevilla, Sevilla, Spain.
Nucleic Acids Res. 2018 Jan 25;46(2):730-747. doi: 10.1093/nar/gkx1240.
The DNA damage response (DDR) is an extensive signaling network that is robustly mobilized by DNA double-strand breaks (DSBs). The primary transducer of the DSB response is the protein kinase, ataxia-telangiectasia, mutated (ATM). Here, we establish nuclear poly(A)-binding protein 1 (PABPN1) as a novel target of ATM and a crucial player in the DSB response. PABPN1 usually functions in regulation of RNA processing and stability. We establish that PABPN1 is recruited to the DDR as a critical regulator of DSB repair. A portion of PABPN1 relocalizes to DSB sites and is phosphorylated on Ser95 in an ATM-dependent manner. PABPN1 depletion sensitizes cells to DSB-inducing agents and prolongs the DSB-induced G2/M cell-cycle arrest, and DSB repair is hampered by PABPN1 depletion or elimination of its phosphorylation site. PABPN1 is required for optimal DSB repair via both nonhomologous end-joining (NHEJ) and homologous recombination repair (HRR), and specifically is essential for efficient DNA-end resection, an initial, key step in HRR. Using mass spectrometry analysis, we capture DNA damage-induced interactions of phospho-PABPN1, including well-established DDR players as well as other RNA metabolizing proteins. Our results uncover a novel ATM-dependent axis in the rapidly growing interface between RNA metabolism and the DDR.
DNA 损伤应答 (DDR) 是一个广泛的信号网络,可被双链 DNA 断裂 (DSBs) 强烈激活。DSB 反应的主要转导蛋白是蛋白激酶共济失调毛细血管扩张突变 (ATM)。在这里,我们确定核多聚(A)结合蛋白 1 (PABPN1) 是 ATM 的一个新靶标,也是 DSB 反应中的关键参与者。PABPN1 通常在调节 RNA 加工和稳定性方面发挥作用。我们确定 PABPN1 作为 DSB 修复的关键调节剂被募集到 DDR 中。一部分 PABPN1 重新定位到 DSB 部位,并以 ATM 依赖的方式在 Ser95 上发生磷酸化。PABPN1 的耗竭使细胞对 DSB 诱导剂敏感,并延长 DSB 诱导的 G2/M 细胞周期阻滞,并且 PABPN1 的耗竭或其磷酸化位点的消除阻碍了 DSB 修复。PABPN1 通过非同源末端连接 (NHEJ) 和同源重组修复 (HRR) 来实现最佳的 DSB 修复,并且特别对于有效的 DNA 末端切除至关重要,这是 HRR 的初始关键步骤。使用质谱分析,我们捕获了磷酸化 PABPN1 的 DNA 损伤诱导的相互作用,包括公认的 DDR 参与者以及其他 RNA 代谢蛋白。我们的结果揭示了在 RNA 代谢和 DDR 之间快速增长的界面中的一个新的 ATM 依赖轴。