Molecular Modulator of RPA and RAD51 in Maintaining Genome Stability

RPA 和 RAD51 维持基因组稳定性的分子调节剂

• 批准号: 10541201
• 负责人: Weihang Chai
• 金额: $ 32.92万
• 依托单位: LOYOLA UNIVERSITY CHICAGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10541201
• 关键词: Affinity; Aging; BRCA mutations; BRCA2 gene; Binding; Binding Proteins; Biochemical; Biological; CRISPR/Cas technology; Cell Cycle Checkpoint; Cells; Cellular Metabolic Process; ChIP-seq; Chromosome Breakage; Complex; DNA; DNA Damage; DNA Repair; DNA biosynthesis; DNA lesion; DNA replication fork; Data; Disease; Environmental Exposure; Equilibrium; Event; Failure; Fiber; Functional disorder; Genetic; Genome; Genome Stability; Genomic Instability; Genomics; Goals; Human; Human Genome; Invaded; Knowledge; Learning; Link; Maintenance; Malignant Neoplasms; Mediating; Molecular; Mutagens; Mutation; PARP inhibition; Pathogenesis; Poly(ADP-ribose) Polymerase Inhibitor; Positioning Attribute; Proteins; Public Health; Rad51 recombinase; Recovery; Repetitive Sequence; Research; Role; SS DNA BP; Single-Stranded DNA; Site; Stress; Surface; System; Techniques; Testing; Therapeutic; Work; anti-cancer therapeutic; cancer type; carcinogenesis; cellular imaging; design; environmental agent; genome integrity; genome-wide; improved; innovation; insight; multidisciplinary; neoplastic cell; nervous system disorder; next generation sequencing; novel; novel strategies; preservation; prevent; protective pathway; protein complex; recruit; repaired; replication stress; response; targeted cancer therapy; tumorigenesis

DNA damage constitutes a major threat to genetic integrity, and has thus been implicated in the pathogenesis of cancer. Elucidating the mechanism safeguarding genome stability is important for understanding the mechanism underlying carcinogenesis. Genome integrity is constantly threatened by endogenous and exogenous agents arising from cellular metabolic processes as well as environmental exposure, many of which impede normal DNA replication and cause replication fork stalling. Stalled forks need to be properly repaired and rescued to prevent DNA lesions and genome instabilities that contribute to tumorigenesis. To repair and rescue stalled replication, a network of proteins regulating DNA damage response, DNA repair, replication, and cell cycle checkpoints are activated in response to stalled replication in order to stabilize and restart stalled forks. However, the mechanism underlying fork repair is poorly understood. Recent studies have revealed that the balance and dynamics of RAD51 and RPA at stalled forks are crucial for fork stabilization and restart. Yet how RAD51 and RPA activities are modulated remains largely elusive. Our recent findings suggest that the high-affinity single-stranded DNA binding protein complex known as CST may be a new modulator for RPA and RAD51 at GC-rich repetitive sequences in response to replication stress. The objective of this proposal is to understand the molecular relationship between CST, RAD51 and RPA in fork rescue, with the goal to provide novel insights into how cells counteract DNA damage caused by genotoxins. We propose to integrate advanced biochemical, cell biological, cell imaging, and next-gen sequencing techniques to examine how CST may regulate RPA binding at stalled sites and maintain fork progression and stability under stress (Aim 1), define how CST may modulate RAD51 activity at GC-rich stalled sites (Aim 2), and characterize the regulatory role of a surface-exposed region of CST in regulating RAD51 and RPA activities (Aim 3). Findings from the proposed research will provide novel information on the mechanism of genome stability maintenance of rescuing stalled replication and preserving genome stability.

DNA 损伤对遗传完整性构成重大威胁，因此与 癌症的发病机制。阐明维护基因组稳定性的机制对于 了解致癌机制。基因组完整性不断受到威胁 细胞代谢过程以及环境因素产生的内源性和外源性因素 暴露，其中许多会阻碍正常的 DNA 复制并导致复制叉停滞。停转的货叉需要 得到适当的修复和拯救，以防止 DNA 损伤和基因组不稳定，从而导致 肿瘤发生。为了修复和挽救停滞的复制，调节 DNA 损伤的蛋白质网络 响应、DNA 修复、复制和细胞周期检查点被激活以响应停滞的复制 为了稳定并重新启动停滞的货叉。然而，人们对分叉修复的机制知之甚少。 最近的研究表明，RAD51 和 RPA 在停滞叉处的平衡和动态对于 货叉稳定并重新启动。然而，RAD51 和 RPA 活动的调节方式在很大程度上仍然难以捉摸。我们的 最近的研究结果表明，称为 CST 的高亲和力单链 DNA 结合蛋白复合物可能 是 RPA 和 RAD51 在富含 GC 的重复序列上响应复制应激的新调节剂。这 该提案的目的是了解 fork 中 CST、RAD51 和 RPA 之间的分子关系 救援，目的是提供关于细胞如何抵抗基因毒素引起的 DNA 损伤的新见解。 我们建议整合先进的生化、细胞生物学、细胞成像和下一代测序 技术来检查 CST 如何调节 RPA 在停滞位点的结合并维持分叉进展，以及 压力下的稳定性（目标 1），定义 CST 如何在富含 GC 的停滞位点调节 RAD51 活性（目标 2）， 并表征 CST 表面暴露区域在调节 RAD51 和 RPA 中的调节作用 活动（目标 3）。拟议研究的结果将为机制提供新的信息 基因组稳定性维持，挽救停滞的复制并保持基因组稳定性。

项目成果

The Intrinsically Disordered Region in the Human STN1 OB-Fold Domain Is Important for Protecting Genome Stability.

• DOI: 10.3390/biology10100977
• 发表时间: 2021-09-28
• 期刊: Biology
• 影响因子: 4.2
• 作者: Chai W;Chastain M;Shiva O;Wang Y
• 通讯作者: Wang Y

Crosstalk between CST and RPA regulates RAD51 activity during replication stress.

• DOI: 10.1038/s41467-021-26624-x
• 发表时间: 2021-11-05
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Lei KH;Yang HL;Chang HY;Yeh HY;Nguyen DD;Lee TY;Lyu X;Chastain M;Chai W;Li HW;Chi P
• 通讯作者: Chi P

CaMKK2 and CHK1 phosphorylate human STN1 in response to replication stress to protect stalled forks from aberrant resection.

• DOI: 10.1038/s41467-023-43685-2
• 发表时间: 2023-11-30
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Jaiswal, Rishi Kumar;Lei, Kai-Hang;Chastain, Megan;Wang, Yuan;Shiva, Olga;Li, Shan;You, Zhongsheng;Chi, Peter;Chai, Weihang
• 通讯作者: Chai, Weihang