Mechanism of Eukaryotic Environmental Mutagenesis

真核环境诱变机制

基本信息

批准号：
10179392
负责人：
GRAHAM C WALKER
金额：
$ 50.72万
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/10179392
关键词：
Adjuvant Chemotherapy Affect Aging Amino Acids Basic Science Binding Biological Biology C-terminal 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 Lead 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 lethal factor anthrax protective factor chemotherapy crosslink environmental chemical environmental mutagens human disease human model improved inhibitor/antagonist innovation insight 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.

项目概要/摘要几乎所有生物体对环境损害做出反应的基本分子机制它们的遗传物质是通过对 DNA 损伤进行跨损伤合成 (TLS) 来获得的。 Rev1/3/7 依赖性突变 TLS 通路对人类健康至关重要，不仅因为它可以通过耐受环境 DNA 损伤（包括修复 DNA 交联）来帮助细胞生存，而且因为这个分子过程是导致绝大多数诱变的原因环境暴露引起的 DNA 损伤会导致癌症、其他人类疾病和 Rev1 是 TLS DNA 聚合酶 Y 家族的成员，而 Rev3 和 Rev7 分别是 TLS DNA 聚合酶 Y 家族的成员。 TLS DNA pol 的催化和非催化亚基这项研究的总体目标是建立在令人兴奋的基础上。 NIEHS 拨款 R01-ES015818 资助的进展，利用人类和哺乳动物生物学，以获得对机制、调节和生理学的详细新见解这一依赖于 Rev1/3/7 的进程在历史上仅在分辨率水平上产生的后果使用具有复杂遗传系统的生物体（例如大肠杆菌和酵母菌）可以实现这项研究的一个特别创新的部分是开发一套新型抑制剂和其他干扰 Rev1/3/7 依赖的 TLS、DNA 交联修复和其他 Rev1/3/7 相关的策略这些不仅是推进生物体如何反应的基础研究的有力探针。来自环境化学物质的 DNA，也有可能改善化疗和其他可能的方法人类健康的一个主要策略是识别抑制 Rev1/3/7 依赖性的小分子。通过干扰通路运行所需的关键相互作用来诱变 TLS，例如 Rev1 100 个氨基酸 C 端结构域 (CTD) 与 DNA pol 的 Rev7 成分的相互作用通过一个接口和其他 TLS DNA 聚合酶的 RIR（Rev1 相互作用区域）通过第二个接口已鉴定出与每个 Rev1 接口结合的示例化合物，并具有预期的生物学效应将在人类肺癌的同源小鼠模型中进行评估。淋巴瘤作为可能的化疗佐剂，可以增加杀伤力，同时减少突变抑制 Rev1/3/7 依赖性诱变 TLS 的其他创新方法包括使用钉合 RIR 肽，使用炭疽保护性抗原将 Rev1 CTD 递送至哺乳动物体内通过将其融合到致死因子的 N 末端来对细胞进行检测，以及测试序列 Rev7 相互作用是否可以发挥作用作为显性负性抑制剂，通过将 Rev7 捕获在非生产性复合物中，一系列影响 Rev1/3/7 依赖性途径中蛋白质的部分功能丧失突变体将产生对复杂蛋白质编排的详细功能见解，这是诱变关键作用的基础 TLS 在 DNA 损伤耐受、交联修复和环境化学物质诱变中的应用。