Cellular responses to DNA replication stress

细胞对 DNA 复制应激的反应

• 批准号: 9316211
• 负责人: Marcus Smolka
• 金额: $ 33.2万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9316211
• 关键词: ATR gene; Aging; Aneuploidy; Animal Model; Apical; BRCA1 Mutation; BRCA1 gene; Binding; Biochemical; Biology; Cancer Patient; Cells; Chromosomal Rearrangement; Complex; DNA; DNA Damage; DNA Repair; DNA Repair Disorder; DNA Repair Pathway; DNA biosynthesis; DNA lesion; DNA replication fork; Europe; Event; Excision; Foundations; Functional disorder; Funding; Genes; Genetic Transcription; Genome; Genomic Instability; Hereditary Breast Carcinoma; Human; Human Cell Line; Individual; Knowledge; Maintenance; Malignant Neoplasms; Malignant neoplasm of ovary; Mammals; Mediating; Modeling; Molecular; Mus; Mutate; Mutation; Nonhomologous DNA End Joining; Orthologous Gene; Outcome; Pathway interactions; Phenotype; Phosphorylation; Phosphotransferases; Play; Predisposition; Proteins; Proteomics; Recruitment Activity; Regulation; Replication Initiation; Replication-Associated Process; Risk; Role; Saccharomycetales; Scaffolding Protein; Signal Transduction; Site; Stress; Testing; Tumor Suppressor Proteins; Work; Yeasts; anticancer research; base; cancer cell; cancer initiation; cancer therapy; design; experimental study; genetic approach; genome editing; homologous recombination; human disease; inhibitor/antagonist; interest; mutant; novel; p53-binding protein 1; prevent; recombinational repair; repaired; response; targeted treatment; tumor progression; tumorigenesis

PROJECT SUMMARY Genomic instabilities drive the progression of cancer, aging, and other human diseases. The integrity of our genome is especially at risk while it is being replicated, as replication forks often encounter obstacles to their progression, including DNA lesions, hard-to-replicate sequences, transcription intermediates, or protein-DNA complexes. These encounters often result in DNA breaks, gross chromosomal rearrangements and aneuploidy, which are key events in cancer initiation. The proper repair of stalled or collapsed replication forks through homologous recombination (HR)-based mechanisms plays a major role in preventing replication stress-induced genomic instabilities and many mutations in components of the HR machinery have been associated with cancer predisposition. In the absence of an intact HR-machinery, error-prone mechanisms such as non-homologous end joining (NHEJ) tend to become hyper-utilized, leading to extensive genomic instability. The regulatory basis of the recruitment of HR and NHEJ factors to DNA lesions is therefore of central interest to genome biology and cancer research, not only for explaining the mechanisms of tumorigenesis, but also for providing promising avenues for cancer therapy, as recently demonstrated for PARP inhibitors that are now approved for treatment of ovarian cancer patients with BRCA1 mutations in Europe and the US. This proposal will investigate a new mechanism for regulation of recombinational DNA repair and repair pathway choice. The central hypothesis is that the evolutionarily conserved scaffolding protein TopBP1 plays a central, yet largely unexplored, role in the control of HR-mediated repair and DNA repair pathway choice. Utilizing biochemical, proteomic and genetic approaches in yeast, human cell lines and genome-edited mice, we will define the molecular mechanism by which TopBP1 and its yeast ortholog Dpb11 control DNA repair. Our studies will provide unparalleled molecular understanding of how the action of key HR and NHEJ factors are coordinated and will reveal how signaling networks integrate the control of DNA replication, checkpoint signaling and DNA repair. Proposed experiments will reveal novel mechanisms of repair pathway choice and recombinational repair that are crucial to suppress genomic instability and cancer. Generated outcomes will have implications in the study of tumorigenesis caused by dysfunctions in HR repair and should provide new rationale for therapy.

项目总结 基因组不稳定性推动癌症、衰老和其他人类疾病的进展。我们的诚信 基因组在被复制时尤其面临风险，因为复制叉经常遇到障碍 进展，包括DNA损伤、难以复制的序列、转录中间产物或蛋白质-DNA 复合体。这些相遇经常导致DNA断裂，总的染色体重排和 非整倍体，这是癌症发生的关键事件。正确修复停滞或倒塌的复制叉子 通过同源重组(HR)的机制在防止复制方面发挥了重要作用 应激诱导的基因组不稳定性和人力资源机制组件的许多突变 与癌症易感性有关。在没有完整的人力资源机制的情况下，容易出错的机制 例如非同源末端连接(NHEJ)倾向于变得高度利用，导致广泛基因组 不稳定。因此，HR和NHEJ因子在DNA损伤中募集的调控基础是 基因组生物学和癌症研究的中心兴趣，不仅是为了解释 肿瘤发生，但也为癌症治疗提供了有希望的途径，最近证明 PARP抑制剂现已被批准用于治疗BRCA1基因突变的卵巢癌患者 欧洲和美国。这项提议将探索一种新的重组DNA调控机制。 修复和修复路径的选择。中心假设是进化上保守的脚手架 TopBP1蛋白在HR介导的修复和DNA调控中发挥着中心作用，但在很大程度上尚未被探索 修复路径选择。利用生化、蛋白质组学和遗传学方法在酵母、人类细胞系和 基因组编辑小鼠，我们将定义TopBP1及其酵母同源基因Dpb11的分子机制 控制DNA修复。我们的研究将提供无与伦比的分子理解，了解关键的HR如何作用 和NHEJ因素是协调的，并将揭示信号网络如何整合DNA的控制 复制、检查点信号和DNA修复。拟议的实验将揭示修复的新机制 途径选择和重组修复是抑制基因组不稳定和癌症的关键。 所产生的结果将对HR修复功能障碍引起的肿瘤发生的研究产生影响 应该为治疗提供新的理论基础。