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.

项目总结（见说明）： 环境损伤可能导致DNA碱基损伤和DNA链断裂，如果不修复，这些结果可能会破坏基因组的稳定性。为了避免基因组不稳定和癌症，DNA聚合酶在准确复制基因组和修复受损DNA方面发挥着核心作用。DNA聚合酶e（Pol E）不仅在复制和修复中发挥重要作用，而且在DNA聚合酶中是独特的，其通过其检查点功能参与协调复制进程和有丝分裂。 Pol e在复制和修复中发挥的重要但尚未精确定义的作用使得对其保真度的研究对于理解人类细胞如何能够避免DNA损伤的后果至关重要，如果不修复，可能导致突变和癌症等疾病。鉴于其重要性，缺乏对人类Pol e保真度的了解与酵母酶保真度的信息量不断增加形成鲜明对比。确定DNA聚合酶对基因组稳定性的贡献的一个重要工具是鉴定和表征降低总体DNA合成保真度的DNA聚合酶的突变衍生物。该项目提出鉴定和表征人类Pol e的突变等位基因， 通过降低复制保真度、改变病变旁路来抑制体内Pol e检查点激活 效率和忠诚度，或者这些活动的某种组合。此外，这些等位基因将用于探测体内复制和修复中的Pole功能。 酵母和人Pol e都已在参与复制的多蛋白复合物中被鉴定， 转录和重组，并已观察到共定位在体内与区域的活性 复制的这些相互作用的特点仍然很差，没有很好地理解，特别是对人类酶。该项目提出通过使用以下方法来确定Pol蛋白相互作用网络： 重组Pol e探针的Pol e相互作用的合作伙伴和特点的功能意义，这些 交互.