Dissecting BRCA1-PALB2 Activity in DNA Repair and Development

剖析 BRCA1-PALB2 在 DNA 修复和发育中的活性

• 批准号: 10388570
• 负责人: Neil Johnson
• 金额: $ 22.9万
• 依托单位: RESEARCH INST OF FOX CHASE CAN CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-05 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10388570
• 关键词: Alleles; Award; BRCA1 gene; Cell Nucleus; Chromatin; Confocal Microscopy; DNA Damage; DNA Double Strand Break; DNA Repair; Development; Equipment; Fanconi' s Anemia; Funding; Goals; Health; Human; Knowledge; Malignant neoplasm of ovary; Mechanics; Microscopy; Mutation; Nanostructures; Nuclear; PALB2 gene; Pathway interactions; Patients; Proteins; Resolution; Site; Time; homologous recombination; improved; insight; malignant breast neoplasm; mutation carrier; parent grant; recruit; repaired

PROJECT SUMMARY/ABSTRACT This submission is for a supplement award for the parent grant R01GM135293: “Dissecting BRCA1-PALB2 activity in DNA repair and development”. The parent grant is currently in its first year, making this the perfect time to upgrade our equipment. The localization of proteins to double stranded DNA breaks (DSBs) can be quantified by the presence of nuclear foci. The number of foci within a nucleus is often indicative of whether a particular protein or repair pathway is functional. A typical focus appears as a homogenous sphere under conventional widefield or confocal microscopy. The information provided does not go beyond whether foci are present or absent. In contrast, super-resolution (SR) microscopy provides high-resolution imagining, which can reveal detailed insights into the DSB-flanking chromatin topology. The ability to perform SR microscopy would greatly enhance the knowledge that can be gained from the current R01-funded project. Not only in understanding the effects of mutations on foci formation, new insights into the mechanics of the DNA damage repair machinery and the interplay between chromatin topology and DNA repair can be revealed. In the parent grant, we aim to uncover the mechanisms by which BRCA1 recruits PALB2 to sites of DNA damage. We proposed to examine the effects of BRCA1 and PALB2 mutations on DNA repair foci. However, SR microscopy has the potential to reveal so much more, beyond simply the presence or absence of foci, such as insights into the nanostructure of BRCA1-PALB2, whether there are separate or overlapping nanodomains, spatial differences, and the BRCA1-PALB2 topology relative to additional homologous recombination (HR) proteins. Moreover, the effects of BRCA1 and PALB2 patient mutations have previously not been examined for their effects on DSB-chromatin topology and nanostructure. The use of SR microscopy will provide super-resolution insight, and ultimately improve our understanding of how BRCA1 and PALB2 mutations impact human health.

项目总结/摘要 本申请是对母基金R 01 GM 135293的补充奖：“解剖BRCA 1-PALB 2 DNA修复和发育”。父母补助金目前是在其第一年，使这是完美的 是时候升级我们的设备了蛋白质在双链DNA断裂（DSB）上的定位可以是 通过核灶的存在来量化。细胞核内病灶的数量通常表明 特定的蛋白质或修复途径是有功能的。一个典型的焦点看起来像一个均匀的球体， 常规的宽视场或共焦显微镜。所提供的信息不超出病灶是否 出席或缺席。相比之下，超分辨率（SR）显微镜提供高分辨率成像，其可以 揭示了对DSB侧翼染色质拓扑结构的详细了解。进行SR显微镜检查的能力将 极大地提高了从当前R 01资助项目中获得的知识。不仅在 理解突变对病灶形成的影响，对DNA损伤机制的新见解 修复机制和染色质拓扑结构和DNA修复之间的相互作用可以揭示。在母 我们的目标是揭示BRCA 1将PALB 2招募到DNA损伤位点的机制。我们 研究BRCA 1和PALB 2突变对DNA修复灶的影响。然而，SR显微镜 有可能揭示更多的东西，不仅仅是焦点的存在或不存在，例如对 BRCA 1-PALB 2纳米结构，无论是分离的还是重叠的纳米结构域，空间 差异，和BRCA 1-PALB 2拓扑结构相对于其他同源重组（HR）蛋白。 此外，BRCA 1和PALB 2患者突变的影响以前没有被检查过。 对DSB-染色质拓扑结构和纳米结构的影响。SR显微镜的使用将提供超分辨率 我们希望通过这项研究，最终提高我们对BRCA 1和PALB 2突变如何影响人类健康的理解。