Mechanisms of Bacterial DNA Polymerase Replication and Fidelity

细菌 DNA 聚合酶复制和保真度的机制

基本信息

批准号：
9273029
负责人：
Janice D Pata
金额：
$ 11.51万
依托单位：
WADSWORTH CENTER
依托单位国家：
美国
项目类别：
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/9273029
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 Health 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 novel therapeutics oxidation pathogen polymerization prevent research study resistant strain response

项目摘要

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而准确的DNA复制的许多机制。即便如此，我们的理解远非完整，尤其是因为现在已知许多不同的聚合酶才能进行完全基因组复制。在这里，我们将研究集中在细菌病原体金黄色葡萄球菌的两种类型的聚合酶上：必需的C-家族复制性聚合酶和Y-家庭translesion聚合酶。 C家族聚合酶包括高度精确的POLC，该POLC无法有效复制受损坏的DNA，以及容易发生的DNAE，该DNAE从RNA引物启动DNA合成。 Y-家庭聚合酶能够复制受损的DNA，但易用错误。我们将使用动力学，结构和分子遗传技术来确定这些聚合酶如何以高精度的高度复制细菌基因组，但也会产生突变，从而引起抗生素反抗。由于DNA复制是这样的基本过程，因此该研究将对其他细菌具有广泛的影响。