Mechanism of Eukaryotic Environmental Mutagenesis

真核环境诱变机制

• 批准号: 7887354
• 负责人: GRAHAM C WALKER
• 金额: $ 37.04万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7887354
• 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; Complex; DNA Damage; DNA-Directed DNA Polymerase; Data; Development; Disease; Escherichia coli; Eukaryota; 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; Play; Polymerase; Positioning Attribute; Process; Proteins; Recruitment Activity; Regulation; Regulator Genes; Research; Research Personnel; Role; 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; 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维修和损害耐受性过程对于保留许多不同代理后的人类健康至关重要。这项研究的长期目标是对真核生物中环境诱变的分子机制进行详细的综合理解。在既复杂又精心控制的分子过程中，当专门的转移合成（TLS）DNA聚合酶复制到由环境药物引起的DNA损伤时，会引入突变。拟议的研究特别强调了Rev1，这既是作为其他TLS DNA聚合酶的脚手架，又是聚合酶本身的脚手架，就在于真核诱变的根源。我们将跟进我们意外的发现，即在G2/m期间，静脉病毒Rev1比G1和S阶段的大多数S. cerevisiae Rev1高出50倍，并通过研究其细胞周期控制的基础，“这主要是在转录后；有希望的候选调节基因，例如UMP1和CDC7等实验，并将确定新的监管基因，并将其进行遗传筛查，并将其进行遗传筛查，并将其引起依次的基因，并将其遗传性基因呈现出来，并将其遗传性基因呈现出来，并将其遗传性基因呈现出来，并将其遗传性基因的范围。对此的控制：使用生化和遗传方法，我们将继续研究Rev1相互作用的结构和功能基础，尤其是Rev1 C末端域，其BRCT域，其BRCT域及其泛素结合基序的相互作用，我们将通过对Rev1 Polymase Action的功能进行了研究。复制特别好。我们将根据REV1-REV7相互作用的破坏开发高通量测定法，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对大多数突变的真核转移综合机制的理解还不足，对基础科学做出了极大的贡献。这些突变有助于衰老，癌症和各种人类疾病。鉴定在这些诱变过程中起作用的其他基因将有助于解决为什么只有某些人在暴露于环境毒素时疾病的问题。 TLS的小分子抑制剂可能导致具有多种应用人类健康的新型“抗氧化剂”药物。