Regulation of DNA replication kinetics by BRCA2 after DNA damage

DNA 损伤后 BRCA2 对 DNA 复制动力学的调节

基本信息

批准号：
10278027
负责人：
Bing Xia
金额：
$ 34.48万
依托单位：
RBHS -CANCER INSTITUTE OF NEW JERSEY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10278027
关键词：
Aftercare Alleles BRCA2 gene Binding Breast Breast Cancer Cell Breast Cancer Model CDC45L gene Cancer Etiology Cell Cycle Checkpoint Cells Chemotherapy and/or radiation Chromatin Cisplatin Complex Coupled DNA DNA Damage DNA Double Strand Break DNA Primase DNA Repair DNA Replication Damage DNA Replication Factor DNA biosynthesis DNA replication fork DNA-Directed DNA Polymerase Defect Development Double Strand Break Repair Family Genes Genome Genome Stability Genomic Instability Germ-Line Mutation Induced Mutation Kinetics Lead MCM10 gene MCM2 gene Malignant Neoplasms Malignant neoplasm of ovary Mammals Mammary Tumorigenesis Maps Mediating Mitotic Modality Mutant Strains Mice Mutation Ovarian Pancreas Pathway interactions Pattern Phenotype Physiological Platinum Play Polymerase Process Prognosis Proteins Radiation Radio Regulation Relapse Replication Error Replication Initiation Replication Origin Reporting Resistance Role S Phase Salts Second Primary Cancers Site Testing Time Travel Tumor Suppression Tumor Suppressor Proteins Xenograft Model base cancer cell cancer therapy conditional knockout design genome integrity helicase homologous recombination inhibitor/antagonist insight member mutant novel strategies overexpression prevent radiation response radioresistant recruit repaired replication stress response skip lesion triple-negative invasive breast carcinoma tumor tumorigenesis

项目摘要

Germline mutations in BRCA2 predispose carriers to breast, ovarian, pancreatic, and other cancers. The gene encodes a very large protein that plays critical roles in genome integrity control by promoting homologous recombination (HR)-mediated repair of DNA double strand breaks (DSBs), DNA damage-induced cell cycle checkpoint, and stability of stalled DNA replication forks, etc. Besides, BRCA2 may play a direct role in DNA replication, as its mutant cells have long been known to have the so-called radio-resistant DNA synthesis (RDS) phenotype, which reflects a defect in the intra-S phase checkpoint, a mechanism that slows down DNA replication after DNA damage presumably to allow time for DNA repair and prevent replication of damage DNA. However, the potential role of BRCA2 in DNA replication initiation or elongation has not been defined. In our preliminary studies, we found that BRCA2 interacts with an essential DNA replication factor, MCM10, and that this interaction restrains replication fork progression, promotes fork stalling and sustains origin firing after DNA damage. We hypothesize that BRCA2 regulates DNA replication kinetics after DNA damage through its interaction with MCM10 and that a dysregulation of replication kinetics after DNA damage, or intra-S phase checkpoint defect, leads to increased mutation rate, cancer development and therapy resistance. We propose 3 specific aims to test the hypotheses. In Aim 1, we will define the mechanisms of BRCA2 function in DNA replication kinetics after DNA damage. In Aim 2, we will determine the role of the BRCA2-MCM10 interaction in DNA replication fidelity and cancer cell response to DNA damaging therapies. In Aim 3, we will determine the role of BRCA2-MCM10 interaction in spontaneous and radiation-induced tumor development. Our findings and proposed studies will elucidate key mechanisms of BRCA2 function in DNA replication, provide new insights into the regulation of replication kinetics after DNA damage, and define the impact of RDS and replication kinetics dysregulation on cancer development and tumor relapse after radiation and chemotherapy.

BRCA2中的生殖线突变使载体易于乳房，卵巢，胰腺和其他癌症。基因编码一种非常大的蛋白质，该蛋白质通过促进同源性在基因组完整性控制中起关键作用重组（HR）介导的DNA双链断裂（DSB）的修复，DNA损伤诱导的细胞周期检查点和停滞的DNA复制叉等的稳定性。此外，BRCA2可能在DNA中起着直接的作用复制，因为长期以来已知其突变细胞具有所谓的抗DNA合成（RDS）表型，它反映了Intra-S相位检查点中的缺陷，这种机制减慢了DNA DNA损伤后的复制大概是为了使DNA修复时间并防止损害的复制脱氧核糖核酸。但是，尚未定义BRCA2在DNA复制启动或伸长率中的潜在作用。在我们的初步研究，我们发现BRCA2与必不可少的DNA复制因子MCM10和这种相互作用限制了复制叉的进展，促进叉子停滞并维持起点发射之后 DNA损伤。我们假设BRCA2通过其DNA损伤后调节DNA复制动力学与MCM10的相互作用以及DNA损伤后复制动力学的失调或Intra s相检查点缺陷，导致突变率提高，癌症发育和耐药性。我们建议 3特定的目的是检验假设。在AIM 1中，我们将定义DNA中BRCA2功能的机制 DNA损伤后的复制动力学。在AIM 2中，我们将确定BRCA2-MCM10相互作用的作用 DNA复制保真度和对DNA损害疗法的癌细胞反应。在AIM 3中，我们将确定 BRCA2-MCM10相互作用在自发和辐射诱导的肿瘤发育中的作用。我们的发现和拟议的研究将阐明BRCA2功能在DNA复制中的关键机制，提供新的见解在DNA损伤后调节复制动力学的调节，并定义RDS的影响和复制的影响放射和化疗后癌症发育和肿瘤复发的动力学失调。