Regulation of polymerase switching in translesion synthesis

跨损伤合成中聚合酶转换的调节

• 批准号: 8609453
• 负责人: Yuichi Machida
• 金额: $ 32.99万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8609453
• 关键词: Base Pairing; Binding; Bypass; Catalytic Domain; Cell Cycle; Cells; Complex; Data; Defect; Deubiquitinating Enzyme; Deubiquitination; Embryo; Environmental Risk Factor; Event; Fibroblasts; Frequencies; Genes; Genetic; Genome; Genome Stability; Genomic Instability; Genotoxic Stress; Goals; Human; Knock-out; Lesion; Light; Maintenance; Malignant Neoplasms; Metabolism; Metalloproteases; Mus; Mutagenesis; Mutation; Nucleotides; Physiological; Play; Polymerase; Prevention; Process; Proteins; Recruitment Activity; Regulation; Role; Site; Slide; Testing; Tumor Suppression; UV induced; Ubiquitin; Ubiquitination; Zinc; dimer; in vitro activity; in vivo; insight; interest; mouse model; novel; prevent; public health relevance; repaired; three-dimensional modeling; tumor

DESCRIPTION (provided by applicant): Environmental factors and endogenous metabolic processes create polymerase-blocking DNA lesions, which can stall DNA replication machinery and cause genome instability. To avoid such events, cells have evolved a mechanism called translesion synthesis (TLS), which uses low fidelity polymerases to allow DNA synthesis across the lesions without actual repair. Our long-term goal is to understand regulatory mechanisms of TLS and determine its importance in prevention of genome instability and tumor formation. Recently, our group and others identified Spartan (also known as DVC1) as a novel regulator of TLS. We demonstrated that Spartan depletion results in accumulation of the error-prone TLS polymerase Pol zeta and higher UV-induced mutagenesis. Building on these observations, we now hypothesize that Spartan negatively regulates Pol zeta complex, and thereby prevents genome instability and tumor formation. To gain more insights into the mechanism of TLS regulation by Spartan and its physiological relevance, we propose to: (1) Study the mechanism of Pol zeta regulation by Spartan, (2) Investigate the cause of DNA damage and cell-cycle defects in Spartan knockout cells, (3) Examine the role of Spartan in genome stability and tumor suppression in mouse. Aim 1 will focus on the SprT domain, a putative metalloprotease domain of Spartan, and determine its role in Pol zeta regulation by characterizing its activity in vitro. n addition, we will investigate how Rev1, a regulator of Pol zeta, is involved in the complex formation of Pol zeta. In Aim 2, we will investigate the cause of DNA damage and cell cycle defects in Spartan knockout cells and determine whether it is related to Spartan's function in TLS. Finally in Aim 3, we will determine whether Spartan is important to prevent mutations, genome instability and tumor formation using mouse models. In summary, these studies will not only shed new light on the regulation of TLS by Spartan, but also provide novel insights into the role of TLS regulation in genome maintenance and tumor suppression.

描述（由申请人提供）：环境因素和内源性代谢过程会产生聚合酶阻断DNA病变，这可以使DNA复制机制停滞不前并引起基因组不稳定性。为了避免这种事件，细胞已经进化出一种称为Translesion合成（TLS）的机制，该机制使用低富达聚合酶允许在不实际修复的情况下允许在整个病变上进行DNA合成。我们的长期目标是了解TLS的调节机制，并确定其对预防基因组不稳定性和肿瘤形成的重要性。最近，我们的小组和其他人确定斯巴达人（也称为DVC1）是TLS的新型调节剂。我们证明，斯巴达耗尽会导致易易见的TLS聚合酶Pol Zeta和较高的紫外线诱导的诱变的积累。在这些观察结果的基础上，我们假设斯巴达人对Pol Zeta复合物进行负调节，从而防止了基因组不稳定性和肿瘤形成。为了更多地了解Spartan及其生理相关性TLS调节的机制，我们建议：（1）研究Spartan通过Spartan进行的POL Zeta调节机制，（2）研究Spartan敲除细胞中DNA损伤的原因和细胞周期缺陷的原因，（3）研究Spartan在Genome stanome stosity stosibal stobilose stabilose andor posity pysibal and tumor中的作用。 AIM 1将重点放在Sprt域，Sprt域是斯巴达坦的假定金属蛋白酶结构域，并通过表征其在体外表征其活性来确定其在Pol Zeta调节中的作用。 n补充，我们将研究Pol Zeta的调节器Rev1如何参与Pol Zeta的复杂形成。在AIM 2中，我们将研究斯巴达敲除细胞中DNA损伤和细胞周期缺陷的原因，并确定它是否与Spartan在TLS中的功能有关。最终，在AIM 3中，我们将确定Spartan对于使用小鼠模型来防止突变，基因组不稳定性和肿瘤形成至关重要。总而言之，这些研究不仅将对斯巴达人的调节进行新的启示，而且还提供了有关TLS调节在基因组维持和抑制肿瘤中的作用的新颖见解。