DNA Polymerase Zeta and Its Role in DNA Damage Tolerance

DNA 聚合酶 Zeta 及其在 DNA 损伤耐受中的作用

• 批准号: 7281582
• 负责人: Gregory N Gan
• 金额: $ 4.37万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7281582
• 关键词: Acrolein; Active Sites; Amino Acids; Antibodies; Benzo(a)pyrene; Binding; Biological Assay; Bypass; Catalytic Domain; Cell Count; Cell Line; Cell Survival; Cells; Chemicals; Chickens; Chromosomal Rearrangement; Cisplatin; Complement; Complementary DNA; Complex; DNA; DNA Damage; DNA Maintenance; DNA biosynthesis; DNA chemical synthesis; DNA lesion; DNA polymerase zeta; DNA-Directed DNA Polymerase; Data; Disruption; Dyes; Embryo; Enzymes; Eukaryota; Eukaryotic Cell; Exclusion; Failure; Fellowship; Fibroblasts; Genome; Genomic Instability; Genomics; Genotoxic Stress; Helix (Snails); Human; Immunoblot Analysis; Individual; Laboratories; Lead; Mammalian Cell; Messenger RNA; Mus; Mutagenesis; Mutagens; Names; Nucleotides; Pattern; Play; Point Mutation; Polymerase; Predisposition; Process; Proteins; Recombinants; Regulation; Resistance; Role; Saccharomyces cerevisiae; Screening procedure; Sequence Homology; Site-Directed Mutagenesis; TP53 gene; Testing; Thinking; Tissues; Transcript; Trypan Blue; Wood material; Yeasts; cytotoxicity; ear helix; environmental agent; enzyme activity; experience; irradiation; luminescence; mutant; pol Gene Products; pol genes; polypeptide; prevent; protein expression; repaired; size; synthetic construct; tumorigenesis

DESCRIPTION (provided by applicant): DNA is constantly being damaged by a variety of cellular and environmental agents. In order to prevent DNA mutagenesis, cells have evolved a variety of effective repair mechanisms to correct damaged DNA. However, during replication, not all DNA damage is repaired, rather, damage instead is tolerated by a group of translesion DNA polymerases such as Polymerase eta and Polymerase zeta (Pol C). Evidence in yeast indicates that the catalytic subunit of Pol delta, Rev3, is capable of extending from misincorporated nucleotide templates and is responsible for both damage-induced and spontaneous mutagenesis. Very little is known about the enzyme in mammalian cells. Data suggests that loss of vertebrate Rev3 leads to increased genomic instability and greater susceptibility to the effects of DNA damage. We plan to study the contribution of Rev3 in damage tolerance when wild-type and Rev3 -/- cells are treated with various DNA damaging agents. Furthermore, we will attempt to determine if the polymerase activity is important in damage bypass and whether truncated portions of the protein is sufficient to restore damage tolerance. Finally, data indicates that Rev3 mRNA is regulated at various levels. Mammalian Rev3 protein expression patterns are unknown. We plan to generate an antibody in order to study tissue specific protein expression and whether Rev3 is upregulated during periods of genotoxic stress via immunoblot analysis.

描述（由申请人提供）：DNA 不断受到各种细胞和环境因素的损害。为了防止DNA突变，细胞进化出了多种有效的修复机制来纠正受损的DNA。然而，在复制过程中，并非所有 DNA 损伤都会得到修复，相反，损伤会被一组跨损伤 DNA 聚合酶（例如聚合酶 eta 和聚合酶 zeta (Pol C)）所耐受。酵母中的证据表明，Pol δ 的催化亚基 Rev3 能够从错误掺入的核苷酸模板延伸，并负责损伤诱导和自发诱变。人们对哺乳动物细胞中的这种酶知之甚少。数据表明，脊椎动物 Rev3 的缺失会导致基因组不稳定性增加，并且更容易受到 DNA 损伤的影响。我们计划研究当野生型和 Rev3 -/- 细胞用各种 DNA 损伤剂处理时 Rev3 在损伤耐受中的贡献。此外，我们将尝试确定聚合酶活性在损伤旁路中是否重要，以及蛋白质的截短部分是否足以恢复损伤耐受性。最后，数据表明 Rev3 mRNA 在不同水平上受到调节。哺乳动物 Rev3 蛋白表达模式尚不清楚。我们计划生成一种抗体，以便通过免疫印迹分析研究组织特异性蛋白表达以及 Rev3 在基因毒性应激期间是否上调。