Mechanism of Eukaryotic Environmental Mutagenesis

真核环境诱变机制

• 批准号: 7477294
• 负责人: GRAHAM C WALKER
• 金额: $ 35.27万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7477294
• 关键词: Active Sites; Address; Affect; Aging; BRCT Domain; Basic Science; Binding; Biochemical Genetics; Biochemistry; Biological; Bypass; C-terminal; CDC7 gene; Cell Cycle; Cell Cycle Regulation; Chromatin; Class; Complex; DNA Damage; DNA-Directed DNA Polymerase; Data; Development; Disease; Disruption; Escherichia coli; Eukaryota; Eukaryotic Cell; Exposure to; Family; Fission Yeast; Genes; Genetic; Genetic Epistasis; Genetic Screening; Goals; Health; Human; Lead; Lesion; Malignant Neoplasms; Molecular; Mutagenesis; Mutation; Nature; Orthologous Gene; Pathologic Mutagenesis; Pharmaceutical Preparations; Phase; Plant Roots; Polymerase; Positioning Attribute; Process; Proteins; Recruitment Activity; Regulation; Regulator Genes; Research; Research Personnel; Role; Role playing therapy; SKP Cullin F-Box Protein Ligases; Saccharomyces cerevisiae; Screening procedure; Testing; Toxic Environmental Substances; Ubiquitin; Ubiquitination; Walkers; adduct; anaphase-promoting complex; base; environmental agent; environmental mutagens; follow-up; genetic regulatory protein; high throughput screening; human disease; inhibitor/antagonist; novel; pol genes; programs; repaired; research study; scaffold; small molecule; tool

DESCRIPTION (provided by applicant): DMA repair and damage tolerance processes are absolutely critical to preserving human health following exposure to many different agents. The long term goal of this research is to develop a detailed integrated understanding of the molecular mechanisms responsible for environmental mutagenesis in eukaryotes. In molecular processes that are both complex and elaborately controlled, mutations are introduced when specialized translesion synthesis (TLS) DNA polymerases copy over DNA damage caused by environmental agents. The proposed research places a special emphasis on Rev1, which by virtue of acting both as a scaffold for other TLS DNA polymerases and as a polymerase itself, lies at the root of eukaryotic mutagenesis. We will follow up on our unanticipated finding that S. cerevisiae Rev1 is expressed 50 fold higher during G2/M than in G1 and most of S phase by investigating the basis of its cell cycle control," which is predominantly posttranscriptional; promising candidate regulatory genes such as UMP1 and CDC7 will be tested, genetic screens for new regulatory genes will be carried out, and experiments will be conducted to determine the importance Of this control: Using both biochemical and genetic approaches, we will continue to investigate the structural and functional basis of Rev1 interactions, focusing particularly on the interactions of the Rev1 C-terminal domain, its BRCT domain, and its ubiquitin-binding motifs. We will investigate the functional importance of the Rev1 polymerase activity by following up on our recent observations suggesting that Rev1 may have a class of cognate lesions it replicates particularly well. We will investigate the function and regulation of S. pombe DinB and its relationship to Rev1. We will develop a high-throughput assay, based on disruption of the Rev1-Rev7 interaction, that will allow screening for inhibitors of eukaryotic environmental mutagenesis. The proposed research will make a highly significant contribution to basic science by elucidating the still poorly understood eukaryotic translesion synthesis mechanisms responsible for most mutations. These mutations contribute to aging, cancer, and various human diseases. Identification of additional genes that play roles in these mutagenic processes will help make it possible to address the question of why only some people develop disease when exposed to an environmental toxin. Small molecule inhibitors of TLS could lead to novel "anti-mutagenesis" drugs with multiple applications to human health.

描述（由申请人提供）：DMA 修复和损伤耐受过程对于在接触多种不同物质后保持人类健康绝对至关重要。这项研究的长期目标是对真核生物环境诱变的分子机制有一个详细的综合理解。在复杂且精心控制的分子过程中，当专门的跨损伤合成 (TLS) DNA 聚合酶复制环境因素引起的 DNA 损伤时，就会引入突变。拟议的研究特别强调 Rev1，它既充当其他 TLS DNA 聚合酶的支架，又充当聚合酶本身，是真核诱变的根源。我们将通过研究其细胞周期控制的基础来跟进我们的意外发现，即酿酒酵母 Rev1 在 G2/M 期间的表达比 G1 和大部分 S 期高 50 倍，”这主要是转录后的；有希望的候选调控基因将测试UMP1和CDC7等新调控基因的遗传筛选，并进行实验以确定这种控制的重要性：使用生化和遗传方法，我们将继续研究结构Rev1 相互作用的功能基础，特别关注 Rev1 C 末端结构域、其 BRCT 结构域及其泛素结合基序的相互作用。我们将通过跟进我们最近的观察结果来研究 Rev1 聚合酶活性的功能重要性。 Rev1 可能具有一类复制得特别好的同源损伤，我们将研究粟酒裂殖酵母 DinB 的功能和调节及其与 Rev1 的关系。我们将开发一种基于 Rev1-Rev7 相互作用破坏的高通量测定，这将允许筛选真核环境诱变的抑制剂。拟议的研究将通过阐明目前仍知之甚少的导致大多数突变的真核跨损伤合成机制，对基础科学做出非常重要的贡献。这些突变会导致衰老、癌症和各种人类疾病。鉴定在这些诱变过程中发挥作用的其他基因将有助于解决为什么只有一些人在暴露于环境毒素时才会患病的问题。 TLS 的小分子抑制剂可能会产生新型“抗突变”药物，对人类健康具有多种应用。