Department of Pharmaceutical Sciences, University of Connecticut, 69 N Eagleville Rd, Unit 3092, Storrs, CT 06269-3092, USA.
Department of Molecular Biology and Biophysics, University of Connecticut Health Center, Farmington, CT 06030, USA.
Structure. 2024 Nov 7;32(11):2134-2146.e3. doi: 10.1016/j.str.2024.09.007. Epub 2024 Oct 3.
REV7 is a HORMA (Hop1, Rev7, Mad2) family adaptor protein best known as an accessory subunit of the translesion synthesis (TLS) DNA polymerase ζ (Polζ). In this role, REV7 binds REV3, the catalytic subunit of Polζ, by locking REV7-binding motifs (RBMs) in REV3 underneath the REV7 safety-belt loop. The same mechanism is used by REV7 to interact with RBMs from other proteins in DNA damage response (DDR) and mitosis. Because of the importance of REV7 for TLS and other DDR pathways, targeting REV7:RBM protein-protein interactions (PPIs) with small molecules has emerged as a strategy to enhance cancer response to genotoxic chemotherapy. To identify druggable pockets at the REV7:RBM interface, we performed computational analyses of REV7 complexed with several RBM partners. The contributions of different interface regions to REV7:RBM stabilization were corroborated experimentally. These studies provide insights into key intermolecular interactions and establish targetable regions of REV7 for the design of REV7:RBM PPI inhibitors.
REV7 是一种 HORMA（Hop1、Rev7、Mad2）家族衔接蛋白，作为跨损伤合成（TLS）DNA 聚合酶 ζ（Polζ）的辅助亚基而广为人知。在这个角色中，REV7 通过将 REV3 下的 REV7 安全带环中的 REV3 结合基序（RBM）锁定，与 Polζ 的催化亚基 REV3 结合。REV7 还使用相同的机制与 DNA 损伤反应（DDR）和有丝分裂中的其他蛋白质的 RBM 相互作用。由于 REV7 对 TLS 和其他 DDR 途径的重要性，使用小分子靶向 REV7:RBM 蛋白-蛋白相互作用（PPIs）已成为增强癌症对遗传毒性化疗反应的一种策略。为了在 REV7:RBM 界面上鉴定可成药的口袋，我们对与几种 RBM 伙伴结合的 REV7 进行了计算分析。不同界面区域对 REV7:RBM 稳定性的贡献通过实验得到了证实。这些研究提供了对关键分子间相互作用的深入了解，并为 REV7:RBM PPI 抑制剂的设计确定了可靶向的 REV7 区域。
