Molecular mechanisms of Werner syndrome helicase in genome stability and aging

维尔纳综合征解旋酶在基因组稳定性和衰老中的分子机制

• 批准号: 10729915
• 负责人: Michael Soniat
• 金额: $ 24.9万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10729915
• 关键词: Acceleration; Aging; Architecture; Award; Binding; Binding Proteins; Biochemical; Biochemistry; Biological; Biological Models; Biology; Biophysics; Bloom Syndrome; Cell Aging; Cell Line; Cells; Cellular biology; Complement; Complex; Cryoelectron Microscopy; DNA; DNA Damage; DNA Repair; DNA Repair Enzymes; DNA Repair Pathway; DNA Structure; Data; Dedications; Disease; Exonuclease; Family; Foundations; Functional disorder; Future; G-Quartets; Genes; Genome; Genome Stability; Genomic Instability; Goals; Hereditary Disease; Heritability; Human; Human Biology; Immune signaling; Individual; Inflammation; Inflammatory; Insecta; Institution; Interferons; Length; Maintenance; Malignant Neoplasms; Mentors; Microscopy; Molecular; Molecular Biology; Monitor; Mutation; Negative Staining; Outcome; PRKDC gene; Pathology; Pathway interactions; Patients; Phase; Phenotype; Play; Positioning Attribute; Premature aging syndrome; Proteins; Repair Complex; Research; Resolution; Role; Signal Transduction; Single-Stranded DNA; Stimulator of Interferon Genes; Structure; Syndrome; TREX1 gene; Testing; Training; Werner Syndrome; Work; age related; biophysical properties; biophysical techniques; career; cytokine; early onset; exodeoxyribonuclease; genome integrity; helicase; insight; interdisciplinary approach; mutant; normal aging; nuclease; overexpression; post-doctoral training; premature; prevent; programs; recruit; replication factor A; senescence; sensor; single molecule; structural biology; telomere; telomere loss; tenure track

PROJECT SUMMARY Progeroid syndromes mimic aging at an accelerated rate and are key to understanding both premature and normal aging. One class of progeroid syndromes results from defective DNA repair pathways. For example, Werner syndrome (WS)—a rare inherited disease characterized by premature aging and cancer—is caused by mutations in the DNA repair helicase WRN. WS patients closely recapitulate many normal aging phenotypes, making WS a model system for aging. However, the cellular and molecular mechanisms involved in WS pathologies remain poorly understood. Here, I propose to answer critical questions regarding the functions of WRN via an interdisciplinary approach that includes structural biology, single-molecule biochemistry, and cell biology. My career goal is to establish an independent research program dedicated to understanding the molecular and cellular mechanisms of genomic instability associated with aging and aging-related diseases. As a first step to achieving this goal, I have pursued postdoctoral training in single-molecule microscopy and biochemistry, complementing my background in structural biology and biophysical techniques. The mentoring phase of the K99/R00 award will provide me with additional training in cryo-electron microscopy, cell biology, telomere biology, and the biology of human aging through an expert group of mentors and advisors. Here I propose to: (1) Determine the molecular architecture of full-length WRN (2) Identify how the nuclease and helicase activities of WRN are regulated (3) Determine how WRN cooperates with telomeric proteins to unwind G-quadruplexes during telomere replication to prevent telomere loss, and (4) determine how WRN-deficiency leads to inflammation and premature cellular senescence. Completion of these aims will represent a major step forward in our understanding of WRN’s role in preventing genomic instability and will lay the groundwork for my long-term goals to determine the mechanisms of genome maintenance by other DNA repair enzymes and their importance in human aging and age-related pathologies. A K99/R00 award will allow me to establish an independent research program that will make me a strong candidate for a tenure-track position at a leading U.S. research institution. My work will provide important insights into how WRN and its interaction partners maintain our genomes and help us understand the biological consequences of WRN dysregulation. Furthermore, these studies will provide a more detailed understanding of how WRN-deficiency leads to accelerated aging phenotypes found in WS.

项目摘要 前代综合征以加速速率模仿衰老，并且是理解早熟的关键 和正常衰老。一类后代综合征是由DNA修复途径缺陷引起的。例如， Werner综合征（WS） - 一种以早衰的特征的罕见遗传性疾病，是由 DNA修复解旋酶WRN中的突变。 WS患者密切概括了许多正常衰老表型， 使WS成为衰老的模型系统。但是，WS中涉及的细胞和分子机制 病理仍然知之甚少。在这里，我建议回答有关功能的关键问题 WRN通过跨学科方法，包括结构生物学，单分子生物化学和细胞 生物学。 我的职业目标是建立一个专门了解的独立研究计划 与衰老和与衰老有关的疾病相关的基因组不稳定性的分子和细胞机制。作为 实现这一目标的第一步，我在单分子显微镜和 生物化学，完成我在结构生物学和生物物理技术方面的背景。心理 K99/R00奖的阶段将为我提供其他关于低温电子显微镜，细胞生物学的培训 端粒生物学，以及人类衰老的生物学通过专家的导师和顾问。我在这里 提示：（1）确定全长WRN的分子结构（2）确定核酸酶和核酸酶和 调节WRN的解旋酶活性（3）确定WRN与远程蛋白的坐标如何放松 端粒复制期间的G四链体以防止端粒损失，（4）确定WRN缺陷的方式 导致炎症和过早的细胞感应。这些目标的完成将代表一个重大步骤 在我们理解WRN在预防基因组不稳定中的作用的过程中，将为我奠定基础 长期目标以确定其他DNA修复酶维持基因组的机制及其 在人衰老和与年龄有关的病理学中的重要性。 K99/R00奖将使我能够建立一个独立的研究计划，这将使我成为强大的 在美国领先的研究机构担任终身职位的候选人。我的工作将提供重要的见解 WRN及其相互作用伙伴如何维持我们的基因组并帮助我们了解生物学 WRN失调的后果。此外，这些研究将提供对 WS中发现WS的衰老表型如何加速了WR。