Mechanisms of damaged DNA replication in eukaryotes

真核生物 DNA 复制受损的机制

• 批准号: 8092859
• 负责人: M. TODD WASHINGTON
• 金额: $ 25.86万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8092859
• 关键词: Address; Affect; Affinity; Binding; Biochemical Genetics; Biological Assay; Biological Models; Cells; Complex; DNA; DNA Damage; DNA Replication Damage; DNA-Directed DNA Polymerase; Dissociation; Enzymes; Equilibrium; Eukaryota; Event; Exposure to; Genetic; Goals; Human; Kinetics; Lesion; Malignant Neoplasms; Measures; Modeling; Molecular Conformation; Mono-S; Mutation; Nucleotides; Polymerase; Prevention; Process; Proliferating Cell Nuclear Antigen; Proteins; Qualifying; Rad30 protein; Reaction; Recruitment Activity; Research; Site; Slide; Testing; Thermodynamics; Thymine Dimers; Ubiquitination; Work; Yeasts; in vivo; insight; novel; protein protein interaction

DESCRIPTION (provided by applicant): The long-term goal of our research is to understand the replication of damaged DNA in eukaryotes at the thermodynamic, kinetic, and structural levels. DNA damage in the template strand blocks replication by classical DNA polymerases. Consequently cells possess a variety of non-classical DNA polymerases that can replace the classical polymerase stalled at sites of DNA damage and can replicate through the damage. Recent kinetic studies and structural studies have provided substantial insights into how these non-classical polymerases differ from classical polymerases and how they are able to accommodate DNA damage. It remains unclear, however, how non-classical polymerases are recruited to sites of DNA damage, how stalled classical polymerases are displaced from sites of DNA damage, and how replication accessory factors promote nucleotide incorporation opposite DNA damage by non-classical polymerases. To address these issues, we propose studies with the following three specific aims: (1) to determine the effect of other protein factors on nucleotide incorporation opposite DNA damage by non-classical polymerases, (2) to determine the mechanism of non-classical polymerase recruitment during translesion synthesis, and (3) to determine the mechanism of classical polymerase displacement during translesion synthesis. These studies will provide a clear understanding of exactly how non-classical polymerases replace classical polymerases at sites of DNA damage and how other protein factors contribute to the replication of damaged DNA. Furthermore, these studies will contribute to our understanding of the origins of mutations and cancers and will provide insight into their prevention.

描述（由申请人提供）：我们研究的长期目标是了解在热力学，动力学和结构水平上真核生物中受损的DNA的复制。模板链中的DNA损伤通过经典的DNA聚合酶阻断复制。因此，细胞具有多种非经典DNA聚合酶，这些聚合酶可以取代在DNA损伤部位停滞的经典聚合酶，并可以通过损伤复制。最近的动力学研究和结构研究提供了有关这些非经典聚合酶与经典聚合酶的不同以及如何适应DNA损伤的实质性见解。然而，尚不清楚如何募集非古典聚合酶到DNA损伤部位，停滞的经典聚合酶如何从DNA损伤部位移动，以及如何促进非经典聚合酶与DNA损伤相反的核苷酸掺入。为了解决这些问题，我们提出了以下三个特定目的的研究：（1）确定其他蛋白质因子对非经典聚合酶掺入核苷酸对相反的DNA损伤的作用，（2）确定在跨性别酶合成过程中非古典聚合酶募集机制的机制，以及（3）确定跨越过程中的跨性别聚合物的机制。这些研究将清楚地了解非古典聚合酶如何替代DNA损伤部位的经典聚合酶以及其他蛋白质因子如何促进受损DNA的复制。此外，这些研究将有助于我们对突变和癌症起源的理解，并将提供对其预防的见解。