Roles of Human DNA Polymearse Epsilon in Mutagenesis and Genome Stability

人类 DNA 聚合酶 Epsilon 在诱变和基因组稳定性中的作用

• 批准号: 8118527
• 负责人: Zachary F Pursell
• 金额: $ 24.88万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 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 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8118527
• 关键词: Alleles; Biochemical; Biological; Bypass; Catalytic Domain; Cell Cycle; Cell Cycle Arrest; Cell Cycle Regulation; Cells; DNA; DNA Damage; DNA Repair; DNA biosynthesis; DNA lesion; DNA strand break; DNA-Directed DNA Polymerase; Disease; Enzymes; Exposure to; Genetic Recombination; Genetic Transcription; Genome; Genome Stability; Genomic Instability; Holoenzymes; Homologous Protein; Human; Human Genome; In Vitro; Instruction; Knowledge; Lead; Left; Lesion; Literature; Malignant Neoplasms; Measures; Mitosis; Mutagenesis; Mutation; N-terminal; Organism; Orthologous Gene; Outcome; Phenotype; Play; Polymerase; Process; Property; Proteins; Pyrimidine Dimers; Recombinants; Role; Signal Transduction; Site; Testing; Therapeutic; Yeasts; base; design; environmental agent; human DNA; human disease; in vivo; mutant; novel; overexpression; pol Gene Products; prevent; protein complex; reconstitution; repaired; response; tool

PROJECT SUMMARY (See instructions): Environmental insults can lead to DNA base damages and DNA strand breaks, outcomes that have the potential to destabilize the genome if left unrepaired. In order to avoid genome instability and cancer, DNA polymerases play a central role in accurately replicating the genome and repairing damaged DNA. DNA Polymerase e (Pol E) specifically plays important roles not only in replication and repair, but is unique among DNA polymerases in its involvement in coordinating replication progression and mitosis through its checkpoint function. The important, yet not precisely defined roles played by Pol e in replication and repair make the study of its fidelity crucial to understanding how human cells are able to avoid the consequences of DNA damage that, if left unrepaired, can lead to mutagenesis and diseases like cancer. Given its importance, the lack of knowledge of the fidelity of human Pol e stands in sharp contrast to the growing amount of information on the fidelity of the yeast enzyme. One important tool to determine the contribution of a DNA polymerase to genome stability is to identify and characterize mutant derivatives of DNA polymerases that decrease overall DNA synthesis fidelity. This project proposes to identify and characterize mutant alleles of human Pol e that suppress Pol e checkpoint activation in vivo through decreased replication fidelity, altered lesion bypass efficiency and fidelity, or some combination of these activities. In addition, these alleles will be used to probe for Pol e function in replication and repair in vivo. Both yeast and human Pol e have been identified in multi-protein complexes involved in replication, transcription and recombination, and have been observed to colocalize in vivo with regions of active replication. These interactions remain poorly characterized and not well understood, especially for the human enzyme. This project proposes to determine the Pol ¿ protein interaction network by using recombinant Pol e to probe for Pol e interaction partners and characterize the functional significance of these interactions.

项目摘要（请参阅说明）： 环境侮辱会导致DNA碱基损伤和DNA链断裂，如果未经修复，则有可能破坏基因组的结果。为了避免基因组不稳定性和癌症，DNA聚合酶在准确复制基因组和修复受损的DNA方面起着核心作用。 DNA聚合酶E（PolE）不仅在复制和修复中发挥了重要作用，而且在DNA聚合酶中独有的作用在其参与协调复制进程和有丝核的参与中是独一无二的。 pol e在复制和维修中所扮演的重要但尚未确切确定的作用，这使得其忠诚度对于了解人类细胞如何避免DNA损伤的后果至关重要，而DNA损伤的后果，如果未经修复，则会导致诱变和诸如癌症之类的疾病。鉴于它的重要性，缺乏对人类政治忠诚度的了解与越来越多的有关酵母酶保真度的信息形成鲜明对比。确定DNA聚合酶对基因组稳定性的贡献的一种重要工具是识别和表征DNA聚合酶的突变衍生物，以降低总体DNA合成保真度。该项目的提议旨在识别和表征人类pol的突变等位基因 通过减少复制保真度，改变病变旁路，在体内抑制pol e检查点激活 效率和保真度，或这些活动的某种组合。此外，这些等位基因将用于在体内复制和修复中探测Pol e功能。 酵母菌和人类pol e均已在涉及复制的多蛋白络合物中鉴定出来， 转录和重组，并已观察到与活性区域进行体内共定位 复制。这些相互作用的特征和不太理解，尤其是对于人类酶。该项目建议通过使用 重组pol e探测相互作用伙伴并表征这些功能意义 互动。