Structural and Mechanistic Studies of DNA Damage Bypass Pathways in Eukaryotes

真核生物 DNA 损伤旁路途径的结构和机制研究

• 批准号: 10551662
• 负责人: M. TODD WASHINGTON
• 金额: $ 38.66万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2027-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10551662
• 关键词: Aging; Biochemical; Biological; Biophysics; Bypass; Cells; Chromosomal Rearrangement; Complex; DNA Damage; DNA biosynthesis; DNA replication fork; Disease; Ensure; Enzymes; Eukaryota; Genome Stability; Genomic Instability; Goals; Maintenance; Malignant Neoplasms; Multiprotein Complexes; Mutation; Pathway interactions; Polymerase; Positioning Attribute; Process; Public Health; Regulation; Research; System; coping; helicase; insight; programs; tool

Project Summary/Abstract DNA damage is a serious threat to genome stability. This is because it interferes with DNA replication leading to mutations and chromosomal rearrangements – the hallmarks of cancer, aging, and other diseases. To ensure genome stability, cells utilize DNA damage bypass pathways to cope with DNA damage during replication. The long-term goal of our research program is to understand how DNA damage bypass is carried out in eukaryotic systems at the structural and mechanistic level. Our research will focus on two damage bypass pathways: translesion synthesis and template switching. Progress in this field has slowed recently because of the challenges associated with studying how the various bypass components assemble into and function within large, dynamic, multi-protein complexes. We have developed the biochemical, biological, biophysical, computational, and structural tools needed to overcome these challenges. This puts us in a unique position to answer many fundamental questions about damage bypass. Our future research plan is organized into three broad projects. First, we will study the regulation of DNA damage bypass. This will be done by determining how bypass complexes are assembled at stalled replication forks and by determining how this assembly is controlled by PCNA-ubiquitylating enzymes. Second, we will study the mechanisms of translesion synthesis. This will be done by determining how the most appropriate non-classical polymerase is chosen to bypass the damage and by determining how each non-classical polymerase accommodates damaged DNA templates. Third, we will study the mechanisms of template switching. This will be done by determining how the remodeling of the replication fork allows for the bypass of DNA damage and by determining how this process is carried out by fork-remodeling DNA helicases. In answering these questions, we will gain important new insights into the maintenance of genome stability.

项目摘要/摘要 DNA损伤是对基因组稳定性的严重威胁。这是因为它会干扰DNA 复制导致突变和染色体重排 - 癌症的标志， 衰老和其他疾病。为了确保基因组稳定性，细胞利用DNA损伤旁路 在复制过程中应对DNA损伤的途径。我们研究的长期目标 程序是了解如何在真核系统中进行DNA损伤旁路 结构和机械水平。我们的研究将集中于两个损坏旁路途径： Translesion合成和模板切换。最近在这个领域的进展放慢了速度 与研究各种旁路组合如何组装到的挑战有关的挑战 并在大型，动态的多蛋白复合物中功能。我们已经开发了 克服所需的生化，生物学，生物物理，计算和结构工具 这些挑战。这使我们处于一个独特的立场来回答许多基本问题 关于伤害旁路。我们未来的研究计划分为三个广泛的项目。首先，我们 将研究DNA损伤旁路的调节。这将通过确定如何绕过来完成 复合物是在停滞的复制叉上组装的，并通过确定该组件的方式 由PCNA-泛素化酶控制。其次，我们将研究 跨性别合成。这将通过确定最合适的非经典性如何来完成 选择聚合酶绕过损害，并确定每个非古典 聚合酶的可容纳DNA模板损坏。第三，我们将研究 模板切换。这将通过确定复制叉的重塑方式来完成 允许绕过DNA损伤，并确定如何通过 叉形型DNA解旋酶。在回答这些问题时，我们将获得重要的新 洞悉基因组稳定性的维持。