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：
8658009
负责人：
PAUL RUSSELL
金额：
$ 34.45万
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 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8658009
关键词：
ATP phosphohydrolase Antibodies Archaea Architecture Ataxia Telangiectasia Ataxia-Telangiectasia-Mutated protein kinase Binding Biochemical Cells Cellular biology Chemotherapy-Oncologic Procedure Chromosomal Instability Complex DNA DNA Binding DNA Damage DNA Double Strand Break DNA Repair Defect Development Disease Eukaryota Excision Family Fission Yeast Foundations Genetic Genetic Polymorphism Goals Growth Homologous Gene Human Immunodeficiency and Cancer Intervention Ionizing radiation Knowledge Link Malignant Neoplasms Meiosis Molecular Molecular Conformation Mutation Nijmegen Breakage Syndrome Nucleotides Outcome Pathway interactions Patients Phenotype Phosphoserine Phosphothreonine Phosphotransferases Play Predisposition Process Proteins Radiation Tolerance Radiation therapy Research Resistance Resolution Roentgen Rays Role Signal Transduction Solutions Specificity Structural Biochemistry Structure Structure-Activity Relationship Synchrotrons Tail Technology Telomere Maintenance Testing Walkers Work Yeasts anticancer research ataxia telangiectasia mutated protein base biophysical techniques cancer radiation therapy design dimer functional hypothalamic amenorrhea genetic regulatory protein homologous recombination human CHEK1 protein human disease inhibitor/antagonist insight member mutant novel strategies nuclease protein complex public health relevance recombinational repair repaired resistance factors response structural biology tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Cancer predispositions in humans and severe DNA damage phenotypes in yeast result from defects in the Mre11-Rad50-Nbs1 (MRN) complex. MRN plays central and essential roles in repairing DNA double-strand breaks (DSBs) during homologous recombination repair as well as acting in meiosis, antibody hypermutation, telomere maintenance, and DNA damage signaling through ATM kinase. Yet, detailed mechanistic insights into these diverse MRN functions remain limited. The Mre11 nuclease complex with the Rad50 ATPase is conserved from archaea to humans and is regulated by Nbs1 in S. pombe and humans. We propose three Specific Aims to advance knowledge of MRN structural biochemistry, conformations, and interactions relevant to DNA damage repair and signaling functions. To accomplish these Aims, we will apply advanced biophysical techniques, including synchrotron solution X-ray scattering and atomic resolution crystal structure technologies in concert with genetic and mutational analyses in yeast. The proposed integrated biophysical and genetic studies will test hypotheses regarding Mre11's role in DNA target specificity and processing, Rad50's role in ATP-induced conformational controls and architectural interactions, and Nbs1's role in modulating Mre11 and Rad50 activities. The expected results will characterize functionally key Mre11, Rad50 and Nbs1 protein-protein and protein-DNA interfaces, conformations, and interaction architectures. Furthermore, the DNA damage sensitivity observed in the absence of any one member of the MRN complex suggests that our results will form a platform to test the utility of inhibitors that increase cellular sensitivity to ionizing radiation and other DNA damaging agents used for cancer radiotherapy and chemotherapy. Overall the results will connect MRN to cellular outcomes and human disease by defining interactions and mechanisms controlling genetic integrity, cancer resistance, radiotherapy resistance, and predispositions to cancer.

描述（由申请人提供）：人类中的癌症易感性和酵母中严重的DNA损伤表型是由MRE11-RAD50-NBS1（MRN）复合物中的缺陷引起的。在同源重组修复过程中，MRN在修复DNA双链断裂（DSB）方面起着中心作用，以及在减数分裂，抗体过度超成熟，端粒维持和通过ATM激酶中的DNA损伤信号传导中的作用。然而，对这些不同MRN功能的详细机械见解仍然有限。与Rad50 ATPase的MRE11核酸酶复合物从古细菌到人类保守，并受到pombe和人类的NBS1的调节。我们提出了三个特定的目标，以提高对与DNA损伤修复和信号传导功能相关的MRN结构生物化学，构象和相互作用的了解。为了实现这些目标，我们将采用先进的生物物理技术，包括同步溶液X射线散射和原子分辨率晶体结构技术，并与酵母中的遗传和突变分析一起。拟议的综合生物物理和遗传研究将检验有关MRE11在DNA目标特异性和加工，Rad50在ATP诱导的构象控制和建筑相互作用中的作用以及NBS1在调节MRE11和RAD50活性中的作用的假设。预期的结果将表征功能上关键的MRE11，RAD50和NBS1蛋白质蛋白质以及蛋白-DNA接口，构象和相互作用体系结构。此外，在没有MRN复合物的任何一个成员的情况下观察到的DNA损伤敏感性表明，我们的结果将形成一个平台，以测试抑制剂的实用性，从而增加细胞对电离辐射的敏感性以及用于癌症放射治疗和化学疗法的其他DNA损害剂。总体而言，结果将通过定义控制遗传完整性，抗癌性，放射治疗耐药性和对癌症的易感性的相互作用和机制，将MRN与细胞结局和人类疾病联系起来。