Mechanism of Eukaryotic Environmental Mutagenesis

真核环境诱变机制

• 批准号: 10626766
• 负责人: GRAHAM C WALKER
• 金额: $ 51.91万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10626766
• 关键词: Adjuvant Chemotherapy; Affect; Aging; Amino Acids; Basic Science; Binding; Biological; Biology; C-terminal; Cell fusion; Cells; Complex; DNA; DNA Damage; DNA Repair; DNA biosynthesis; DNA crosslink; DNA lesion; DNA-Directed DNA Polymerase; Development; Dominant-Negative Mutation; Environmental Exposure; Escherichia coli; Family; Funding; Genetic; Genetic Diseases; Genetic Materials; Goals; Grant; Health; Human; Human Genetics; Lung Lymphoma; Malignant Neoplasms; Malignant neoplasm of lung; Mammalian Cell; Molecular; Mutagenesis; Mutation; National Institute of Environmental Health Sciences; Organism; Pathway interactions; Peptides; Physiological; Process; Proteins; Regulation; Research; Resistance; Resolution; Role; Saccharomyces cerevisiae; Series; System; Testing; Therapeutic; anthrax protective factor; chemotherapy; crosslink; environmental chemical; environmental mutagens; human disease; human model; improved; inhibitor; innovation; insight; lethal factor; loss of function; member; mouse model; mutant; novel; novel drug class; operation; protein complex; repaired; response; small molecule; virtual

Project Summary/Abstract A fundamental molecular mechanism by which virtually all organisms respond to environmental damage to their genetic material is by carrying out translesion synthesis (TLS) over DNA lesions. The eukaryotic Rev1/3/7-dependent pathway of mutagenic TLS is critically important to human health, not only because it can help cells to survive by tolerating environmental DNA damage, including the repair of DNA crosslinks, but also because this molecular process is responsible for the vast majority of the mutagenesis that occurs as a result of damage to DNA. Mutations from environmental exposure contribute to cancer, other human diseases, and aging. Rev1 is member of the Y family of TLS DNA polymerases, while Rev3 and Rev7 respectively are the catalytic and non-catalytic subunits of TLS DNA pol ζ. The overall goal of this research is to build on exciting progress funded by NIEHS grant R01-ES015818 by taking advantage of new developments in human and mammalian biology to gain detailed new insights into the mechanism, regulation, and physiological consequences of this Rev1/3/7-dependent process at a level of resolution that has historically only been attainable using organisms with sophisticated genetic systems such as Escherichia coli and Saccharomyces cerevisiae. A particularly innovative component of this research is to develop a suite of novel inhibitors and other strategies to interfere with Rev1/3/7-dependent TLS, DNA crosslink repair, and other Rev1/3/7-related processes. These will not only be powerful probes to advance basic research into how organisms respond to DNA from environmental chemicals, but also have the potential to improve chemotherapy and possibly other aspects of human health. One major strategy is to identify small molecules that inhibit Rev1/3/7-dependent mutagenic TLS by interfering with critical interactions required for operation of the pathway, such as the interaction of the Rev1 100 amino acid C-terminal domain (CTD) with the Rev7 component of DNA pol ζ through one interface and the RIR (Rev1-interacting region) of other TLS DNA polymerases through a second interface. Exemplar compounds have already been identified that bind to each Rev1 Interface and have the expected biological effects. These will be evaluated in syngeic mouse models of human lung cancer and lymphoma as possible chemotherapy adjuvants that increase killing while also reducing the mutagenesis that gives rise to resistance. Other innovative approaches to inhibiting Rev1/3/7-dependent mutagenic TLS include using stapled RIR peptides, using the Anthrax Protective Antigen to deliver the Rev1 CTD into mammalian cells by fusing it to the N-terminus of Lethal Factor, and testing whether Rev7-interacting sequences can serve as dominant negative inhibitors by trapping Rev7 in nonproductive complexes. In a complementary approach, a series of partial-loss-of-function mutants affecting proteins in the Rev1/3/7-dependent-pathway will yield detailed functional insights into the complex protein choreography that underlies the crucial roles of mutagenic TLS in DNA damage tolerance, crosslink repair, and mutagenesis from environmental chemicals.

项目摘要/摘要 一种基本的分子机制，几乎所有的有机体都通过它来应对环境破坏 他们的遗传物质是通过在DNA损伤上进行跨损伤合成(TLS)。真核生物 依赖Rev1/3/7的突变TLS途径对人类健康至关重要，不仅是因为它可以 通过容忍环境中的DNA损伤来帮助细胞存活，包括修复DNA交联链，但也 因为这一分子过程导致了绝大多数的诱变 对DNA的损害。暴露在环境中的突变会导致癌症、其他人类疾病以及 衰老。Rev1是TLS DNA聚合酶Y家族的成员，而Rev3和Rev7分别是 TLS DNA Pollζ的催化和非催化亚基。这项研究的总体目标是在激动人心的基础上 由NIEHS赠款R01-ES015818资助的进展，利用人类和 哺乳动物生物学将获得对机制、调节和生理的详细新见解 这一依赖于Rev1/3/7的进程在历史上只有 可以使用具有复杂遗传系统的生物，如大肠杆菌和酵母菌 酿酒。这项研究的一个特别创新的组成部分是开发一套新型抑制剂和 干扰依赖Rev1/3/7的TLS、DNA交联链修复和其他Rev1/3/7相关的其他策略 流程。这些不仅将是推进生物体如何响应的基础研究的强大探测器 来自环境化学品的DNA，但也有可能改善化疗和其他 人类健康的各个方面。一种主要的策略是识别抑制Rev1/3/7依赖的小分子 通过干扰该途径操作所需的关键相互作用，如 DNAPollζ中Rev1100氨基酸C端区与Rev7组分的相互作用 通过一个界面和其他TLS DNA聚合酶的RIR(Rev1相互作用区)通过第二个界面 界面。已经确定了与每个Rev1接口结合并具有 预期的生物效应。这些将在人类肺癌和肺癌的综合小鼠模型中进行评估。 淋巴瘤作为可能的化疗辅助剂，在增加杀伤力的同时也减少了 会引起抵抗。抑制Rev1/3/7依赖突变TLS的其他创新方法包括 利用装订的RIR多肽，利用炭疽保护抗原将Rev1 CTD导入哺乳动物体内 通过将其与致死因子的N端融合，并测试Rev7相互作用的序列是否可以服务于 作为主要的负性抑制物，通过将Rev7捕获在非生产性复合体中。在一种补充方法中， 一系列影响Rev1/3/7依赖途径中蛋白质的部分功能丧失的突变体将产生 对复杂蛋白质编排的详细功能洞察，这是诱变剂关键作用的基础 TLS在环境化学品的DNA损伤耐受性、交联链修复和诱变方面的作用。

项目成果

Evolution of Rev7 interactions in eukaryotic TLS DNA polymerase Polζ.

• DOI: 10.1016/j.jbc.2022.102859
• 发表时间: 2023-03
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: McPherson, Kerry Silva;Rizzo, Alessandro A.;Erlandsen, Heidi;Chatterjee, Nimrat;Walker, Graham C.;Korzhnev, Dmitry M.
• 通讯作者: Korzhnev, Dmitry M.

A stapled POL κ peptide targets REV1 to inhibit mutagenic translesion synthesis.

钉钉的polκ肽靶向Rev1以抑制诱变转移合成。

• DOI: 10.1002/em.22395
• 发表时间: 2020-10
• 期刊: Environmental and molecular mutagenesis
• 影响因子: 2.8
• 作者: Chatterjee N;D'Souza S;Shabab M;Harris CA;Hilinski GJ;Verdine GL;Walker GC
• 通讯作者: Walker GC