Mre11/Rad50/Nbs1 Structural Biology for DNA Damage Responses

Mre11/Rad50/Nbs1 DNA 损伤反应的结构生物学

基本信息

批准号：
8888891
负责人：
PAUL RUSSELL
金额：
$ 40.51万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-08-01 至 2020-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8888891
关键词：
ATM Gene Mutation ATP phosphohydrolase Address Advanced Development Advanced Malignant Neoplasm Antibodies Ataxia Telangiectasia Ataxia-Telangiectasia-Mutated protein kinase Binding Biochemical Biochemical Genetics Biochemistry Biological Biological Assay Biology Cancer Etiology Cell Death Cells Cellular biology Chemicals Chromosomal Instability Chromosomes Complex Coupled DNA DNA Binding DNA Damage DNA Double Strand Break DNA Integration DNA Repair DNA Repair Pathway Defect Development Disease Eukaryota Fission Yeast Funding Future Genetic Genomic Instability Human Hydrolysis Immunodeficiency and Cancer Inherited Knowledge Lead Link Malignant Neoplasms Mediating Medicine Meiosis Molecular Molecular Conformation Mutagenesis Mutate Mutation Neurodegenerative Disorders Nijmegen Breakage Syndrome Nonhomologous DNA End Joining Normal Cell Outcome Pathway interactions Patients Phenotype Play Predisposition Process Protein Binding Proteins Radiation Radiation Tolerance Recruitment Activity Research Project Grants Resistance Resolution Roentgen Rays Role Signal Pathway Signal Transduction Solutions Structural Biochemistry Structure Structure-Activity Relationship Technology Telomere Maintenance Testing Therapeutic Intervention United States National Institutes of Health Work Yeasts ataxia telangiectasia mutated protein base cancer cell cancer therapy chemotherapy design endoexonuclease endonuclease genetic analysis human disease inhibitor/antagonist insight killings microbial mutant nuclease public health relevance recombinational repair repaired response small molecule structural biology success telomere tool yeast genetics

项目摘要

DESCRIPTION (provided by applicant): Defects in the Mre11-Rad50-Nbs1 (MRN) complex results in human disease including cancer, and severe DNA damage phenotypes in yeast. MRN, acting with CtIP, is essential for the repair of double- stranded DNA breaks (DSBs) by homologous recombinational repair (HRR), as well as acting in meiosis, antibody hypermutation, telomere maintenance, rescue of stalled replication forks, and DNA damage signaling through ATM kinase. Yet, the mechanistic basis for diverse MRN functions is poorly understood. We propose three Specific Aims to understand MRN, CtIP and ATM structural biochemistry, activities, conformations and interactions relevant for DSB repair and signaling. We will couple advanced biophysical technologies, including atomic-resolution crystal structures and small- angle X-ray scattering in solution, with mutagenesis, biochemistry and yeast genetic analyses. Our integrated approaches will test hypotheses that dynamic MRN conformations and macromolecular interfaces control biological responses at DSBs. In particular, our results will significantly advance knowledge of 1) Mre11 DNA-binding and nuclease mechanisms and their importance for DNA repair pathway choice and progression. 2) How Rad50 binds to DNA and uses its ATPase activity to both handoff DNA to Mre11 and allosterically regulate Mre11 nuclease activities. 3) A Rad50 patient mutation that will advance our understanding of therapeutically targetable Rad50 protein features. 4) Catalytic and non-catalytic roles of CtIP. 5) How MRN recruits and activates ATM at DSBs. Our latest Mre11 inhibitor results and work from others in the field suggest that MRN roles in DSB repair and signaling are viable targets for the development of advanced adjunct cancer therapies, which work by synthetic lethality with current radiation and chemotherapies along with weaknesses in other DNA repair or signaling pathways arising from either inhibitors or cancer-specific genetic defects. Thus, our integrated results will provide a molecular framework for understanding cancer etiologies from DNA repair defects and for the design of advanced cancer therapies targeted against specific MRN activities. Collectively, project results will connect MRN, CtIP and ATM to cellular outcomes and human disease-states by defining interactions, conformations and mechanisms critical for genetic integrity, cancer therapy resistance, and future cancer treatments.

描述（由申请人提供）：Mre11-Rad50-Nbs1 (MRN) 复合物的缺陷会导致包括癌症在内的人类疾病，而酵母中的严重 DNA 损伤表型与 CtIP 一起作用，对于双链 DNA 的修复至关重要。通过同源重组修复 (HRR) 断裂 (DSB)，以及在减数分裂、抗体超突变、端粒维持、复制停滞的挽救中发挥作用然而，我们对不同 MRN 功能的机制基础知之甚少，以了解与 DSB 修复和信号传导相关的 MRN、CtIP 和 ATM 结构生物化学、活性、构象和相互作用。我们将先进的生物物理技术（包括原子分辨率晶体结构和溶液中的小角度 X 射线散射）与诱变、生物化学和酵母遗传分析相结合，我们的综合方法将测试动态 MRN 构象的假设。特别是，我们的结果将显着增进对以下方面的认识：1) Mre11 DNA 结合和核酸酶机制及其对 DNA 修复途径选择和进展的重要性；2) Rad50 如何与 DNA 结合并使用其 ATP 酶。 3) Rad50 患者突变将增进我们对治疗靶向 Rad50 蛋白的理解。 4) CtIP 的催化和非催化作用。 MRN 如何在 DSB 中招募和激活 ATM 我们最新的 Mre11 抑制剂结果以及该领域其他人的工作表明，MRN 在 DSB 修复和信号传导中的作用是开发先进辅助癌症疗法的可行目标，这些疗法通过当前辐射的合成致死作用发挥作用。因此，我们的综合结果将为了解 DNA 修复缺陷的癌症病因和晚期癌症的设计提供分子框架。总的来说，项目结果将通过定义对遗传完整性、癌症治疗耐药性和未来癌症治疗至关重要的相互作用、构象和机制，将 MRN、CtIP 和 ATM 与细胞结果和人类疾病状态联系起来。