Mechanism of Eukaryotic Environmental Mutagenesis

真核环境诱变机制

基本信息

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
项目类别：
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

项目摘要

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，几乎所有生物都会响应环境损害对环境的损害它们的遗传物质是通过在DNA病变上进行跨性别合成（TLS）。真核生物诱变TLS的Rev1/3/7依赖性途径对人类健康至关重要，不仅是因为它可以通过耐受环境DNA损伤，包括DNA交联的修复，还可以帮助细胞生存因为这种分子过程是导致绝大多数发生的诱变。对DNA的损害。环境暴露的突变会导致癌症，其他人类疾病和老化。 Rev1是TLS DNA聚合酶Y家族的成员，而Rev3和Rev7分别是 TLS DNApolζ的催化和非催化亚基。这项研究的总体目标是基于激动人心由NIEHS资助R01-ES015818资助的进度，利用人类和哺乳动物生物学，以获取有关机制，调节和生理学的新见解这种Rev1/3/7依赖性过程的后果是在历史上仅存在的解决方案级别可以使用有机体与复杂的遗传系统（例如大肠杆菌和糖疗法）一起获得酿酒酵母。这项研究的一个特别创新的组成部分是开发一套新型的抑制剂和干扰Rev1/3/7依赖性TL的其他策略，DNA交叉链接维修和其他Rev1/3/7 过程。这些不仅将是对生物如何反应的基础研究的强大探针来自环境化学物质的DNA，但也有可能改善化学疗法和其他可能人类健康方面。一种主要策略是确定抑制Rev1/3/7依赖性的小分子诱变TLS通过干扰途径运行所需的关键相互作用，例如 Rev1 100氨基酸C末端结构域（CTD）与DNApolζ的Rev7分量的相互作用通过一个界面和其他TLS DNA聚合酶的RIR（Rev1相互作用区域）通过第二个界面。已经确定了符合每个Rev1接口的示例化合物，并具有预期的生物学效应。这些将在人类肺癌的合成小鼠模型和淋巴瘤作为可能增加杀伤性的化学疗法调节剂，同时还减少了诱变产生抵抗。抑制Rev1/3/7依赖性诱变TL的其他创新方法包括使用分阶段的RIR肽，使用炭疽保护抗原将Rev1 CTD输送到哺乳动物中通过将其融合到致命因子的N末端，并测试Rev7相互作用序列是否可以使用通过将Rev7捕获在非生产性复合物中，作为主要的负抑制剂。在完整的方法中一系列影响Rev1/3/7依赖性pathway中蛋白质的功能突变体将产生对复杂蛋白编排的详细功能见解，该编排是诱变的关键作用的基础 DNA损伤耐受性，交联修复和环境化学化学诱变中的TLS。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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.