Mechanisms of Bacterial DNA Polymerase Replication and Fidelity

细菌 DNA 聚合酶复制和保真度的机制

• 批准号: 8817982
• 负责人: Janice D Pata
• 金额: $ 28.49万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8817982
• 关键词: Anti-Bacterial Agents; Antibiotic Resistance; Antibiotics; Bacteria; Bacterial DNA; Bacterial Genome; Bacterial Infections; Biochemical; Cells; Centers for Disease Control and Prevention (U.S.); Cessation of life; Ciprofloxacin; DNA; DNA Damage; DNA Double Strand Break; DNA Replication Damage; DNA Structure; DNA biosynthesis; DNA-Directed DNA Polymerase; Data; Development; Drug resistance; Enzymes; Exonuclease; Family; Foundations; Frequencies; Funding; Genetic Techniques; Genome; Genome Stability; Gram-Positive Bacteria; Grant; Guanine + Cytosine Composition; Horizontal Gene Transfer; Hospitals; Human; In Vitro; Induced Mutation; Infection; Kinetics; Methicillin Resistance; Molecular Genetics; Movement; Mutation; Nucleotides; Polymerase; Positioning Attribute; Process; Proteins; RNA primers; Reporting; Research; Role; Running; SOS Response; Source; Specificity; Staphylococcus aureus; Structure; Work; bactericide; combat; cytotoxic; cytotoxicity; defined contribution; design; genetic approach; in vivo; inhibitor/antagonist; methicillin resistant Staphylococcus aureus; novel; oxidation; pathogen; polymerization; prevent; public health relevance; research study; resistant strain; response

DESCRIPTION (provided by applicant): Since the discovery of the double-helical structure of DNA 60 years ago, a wealth of kinetic and structural data have illuminated many of the mechanisms DNA polymerases use to achieve efficient and accurate DNA replication. Even so, our understanding is far from complete, especially since many different polymerases are now known to be required for complete genome duplication. Here, we focus our research on two types of polymerases from the bacterial pathogen Staphylococcus aureus: the essential C-family replicative polymerases and the Y-family translesion polymerases. The C-family polymerases include the highly accurate PolC, which is unable to efficiently copy damaged DNA, and the error-prone DnaE, which initiates DNA synthesis from RNA primers. The Y-family polymerases are able to replicate damaged DNA but are highly error-prone. We will use kinetic, structural and molecular genetic techniques to determine how these polymerases replicate the bacterial genome with high accuracy overall, but also create mutations that give rise to antibiotic resistance. Since DNA replication is such a fundamental process, the research will have broad implications for other bacteria.

描述（由申请人提供）：自从60年前发现DNA的双螺旋结构以来，大量的动力学和结构数据已经阐明了DNA聚合酶用于实现有效和准确的DNA复制的许多机制。即便如此，我们的理解还远未完成，特别是因为现在已知许多不同的聚合酶是完整基因组复制所必需的。在这里，我们把我们的研究集中在两种类型的聚合酶从细菌病原体金黄色葡萄球菌：必要的C-家族复制型聚合酶和Y-家族translesion聚合酶。C家族聚合酶包括高度精确的PolC，它不能有效地复制受损的DNA，和易错的DnaE，它从RNA引物开始DNA合成。Y家族聚合酶能够复制受损的DNA，但高度易错。我们将使用动力学，结构和分子遗传学技术来确定这些聚合酶如何以高精度复制细菌基因组，同时也产生抗生素的突变。 阻力由于DNA复制是一个如此基本的过程，这项研究将对其他细菌产生广泛的影响。