Structure/Function Studies of DNA Replication Initiation

DNA复制起始的结构/功能研究

• 批准号: 8123707
• 负责人: JAMES M BERGER
• 金额: $ 7万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8123707
• 关键词: ATP phosphohydrolase; ATPase Domain; Area; Award; Binding; Biochemical; Biological Assay; Biological Factors; Cells; Chemicals; Chromosomes; Complex; DNA; DNA Binding; DNA Primase; DNA replication origin; DNA-Directed DNA Polymerase; Deposition; DnaB helicase; DnaG Protein; Enzymes; Escherichia coli; Event; Family; Foundations; Genetic Materials; Genomic Instability; Goals; Hand; Helix-Turn-Helix Motifs; Homo; Image; Investigation; Knowledge; Lead; Life; Link; Malignant Neoplasms; Methods; Modeling; Molecular; Molecular Structure; Mutagenesis; Neoplasms; Nucleic Acids; Nucleotides; Oligonucleotides; Peptide Initiation Factors; Polymerase; Positioning Attribute; Process; Prokaryotic Cells; Proteins; Recruitment Activity; Replication Initiation; Replication Origin; Research; Single-Stranded DNA; Site; Structure; Structure-Activity Relationship; Testing; Work; base; cell transformation; crosslink; drug development; ds-DNA; helicase; inhibitor/antagonist; melting; novel; paralogous gene; public health relevance; response; scaffold; small molecule

DESCRIPTION (provided by applicant): The faithful propagation of genetic material is essential to life. A central prerequisite to the copying of DNA is the dedicated assembly of replicative machineries at proper sites on the chromosome. This replication "initiation" event depends on multiple components, including: 1) initiators, ATPases that bind origins and recruit other proteins to the replication start site, 2) helicase-loaders, which deposit replicative helicases onto DNA, and 3) primases, enzymes that create short oligonucleotides for extension by DNA polymerases. A long-term objective of our research has been to understand the molecular structure/function relationships that govern the initiation of DNA replication. Many of the proteins responsible for initiation have been identified, and a preliminary framework for their action is in place. However, there remains a host of outstanding questions regarding how these factors act individually and cooperatively to construct a competent replication fork. Our prior efforts in this area have generated new mechanistic models for initiator action, helicase loading, and priming that we are now in an ideal position to test. Using a combination of structural, biochemical, and biophysical methods we aim to: 1) Define how ATP controls DNA binding and remodeling by prokaryotic initiators, 2) Determine how ATP regulates bacterial helicase-loader assembly and function, and 3) Establish how the bacterial primase synthesizes primers and is inhibited by disparate types of small-molecule agents, including the stringent response regulator, (p)ppGpp. Together, these efforts will impact a number of scientific fronts, from understanding how origins are engaged and restructured to promote replisome construction, to defining how chemical energy and small-molecule inhibitors control the function of dynamic macromolecular machines. Knowledge of these processes is necessary for understanding how cells ultimately avoid initiation errors linked to genomic instabilities, transformation, and neoplasia. PUBLIC HEALTH RELEVANCE: The faithful propagation of genetic material is essential to life. The goal of our research is understand the structure/function relationships that govern the initiation of DNA replication at a molecular level. Knowledge of these events is necessary for understanding how cells ultimately avoid errors linked to genomic instabilities and cancer onset.

描述（由申请人提供）：遗传物质的忠实传播对生命至关重要。复制DNA的主要先决条件是在染色体上适当的位置的复制机械组装。此复制“启动”事件取决于多个组件，包括：1）启动器，结合起源并募集其他蛋白质与复制起始位点的ATPase，2）解旋酶载荷器，它们将复制性解旋酶沉积到DNA和3）起源酶，以及在DNA polymassase extrymass assels extrymassass extrymasase assemens asspressions extrymasases shore primase sopimass酶。我们研究的一个长期目标是了解控制DNA复制启动的分子结构/功能关系。已经确定了许多负责启动的蛋白质，并且采取了行动的初步框架。但是，关于这些因素如何单独和合作以构建有能力的复制叉的方式，仍然存在许多杰出问题。我们在该领域的先前努力为引发器动作，解旋酶负载和启动启动生成了新的机械模型，我们现在处于测试的理想位置。使用结构，生化和生物物理方法的结合，我们的目的是：1）定义ATP如何控制DNA如何控制DNA的结合和重塑，2）确定ATP如何调节细菌量载体载荷量的组装组装和功能，以及3）建立细菌原始响应如何通过小细胞的响应来确定差异化和差异，并通过差异化的类型，是差异化的，差异化。调节器，（p）ppgpp。从了解如何参与和重组来促进重新构体结构到定义化学能和小分子抑制剂如何控制动态大分子机器的功能，从了解原产地参与并进行了重组，共同影响了许多科学方面。这些过程的知识对于理解细胞最终如何避免与基因组不稳定性，转化和肿瘤有关的启动错误所必需。公共卫生相关性：忠实的遗传材料传播对生活至关重要。我们研究的目的是了解控制DNA复制在分子层的启动的结构/功能关系。对这些事件的了解对于理解细胞最终如何避免与基因组不稳定性和癌症发作有关的错误是必要的。