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：
9055648
负责人：
PAUL RUSSELL
金额：
$ 41.15万
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/9055648
关键词：
ATM Gene Mutation ATP phosphohydrolase Address Advanced Development Advanced Malignant Neoplasm Antibodies Binding Binding Proteins 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 DNA replication fork Defect Development Eukaryota Fission Yeast Funding Future Genetic Genomic Instability Health 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 Proteins Radiation Radiation Tolerance Recruitment Activity Research Project Grants Resistance Resolution Roentgen Rays Role Signal Pathway Signal Transduction Structural Biochemistry Structure Structure-Activity Relationship Technology Telomere Maintenance Testing Therapeutic Intervention United States National Institutes of Health Work Yeasts ataxia telangiectasia mutated protein ataxia-telangiectasia like disorder base cancer cell cancer therapy chemotherapy design endoexonuclease endonuclease genetic analysis human disease inhibitor/antagonist insight killings microbial mutant nuclease 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（由适用提供）：MRE11-RAD50-NBS1（MRN）复合物中的缺陷导致人类疾病，包括癌症，以及酵母中严重的DNA损伤表型。与CTIP作用的MRN对于通过同源重组修复（HRR）（HRR）以及在减数分裂，抗体超突变，端粒维持，维持端粒维持，挽救停滞的复制叉和通过ATM KINASE的DNA损伤信号中的作用是通过同源重组修复（HRR）修复双链DNA断裂（DSB）至关重要的。然而，潜水员MRN功能的机械基础知之甚少。我们提出了三个特定的目标，以了解与DSB修复和信号相关的MRN，CTIP和ATM结构生物化学，活动，构象和相互作用。我们将与溶液中的原子分辨率晶体结构和小角度X射线散射在内，以及诱变，生物化学和酵母遗传分析。我们的综合方法将检验假设，即动态MRN构象和大分子界面控制DSB的生物学反应。特别是，我们的结果将显着提高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结合并利用其ATPase活性将DNA转换为MRE11，并通过变构调节MRE11核酸酶活性。 3）RAD50患者突变，该突变将提高我们对热靶向Rad50蛋白特征的理解。 4）CTIP的催化和非催化作用。 5） MRN如何招募和激活DSB的ATM。我们最新的MRE11抑制剂结果和该领域其他人的工作表明，DSB修复和信号传导中的MRN角色是开发晚期辅助癌症疗法的可行靶标，这些目标是通过合成的杀伤力与当前的辐射和化学疗法以及其他DNA修复中的弱点以及来自其他DNA修复的弱点或来自其他DNA修复的弱点，或者来自其他抑制剂或癌症遗传性遗传的遗传。这是我们的综合结果将为了解DNA修复缺陷的癌症病因以及针对特定MRN活性的晚期癌症疗法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，抗癌治疗耐药性和未来癌症治疗至关重要的相互作用，会议和机制，将MRN，CTIP和ATM与细胞结局和人类疾病状态联系起来。