Regulatory circuitry controlling the cell cycle of Helicobacter pylori

控制幽门螺杆菌细胞周期的调节电路

• 批准号: 9207987
• 负责人: Paola E Mera
• 金额: $ 14.8万
• 依托单位: NEW MEXICO STATE UNIVERSITY LAS CRUCES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9207987
• 关键词: ATP phosphohydrolase; Antibiotics; Bacteria; Binding; Binding Proteins; Biological Assay; Biological Models; Cancer Etiology; Caulobacter crescentus; Cell Cycle; Cell Cycle Progression; Cell Proliferation; Cell Separation; Cells; Cessation of life; ChIP-seq; Chromosome Segregation; Chromosomes; Cytokinesis; DNA; DNA biosynthesis; Data; Development; Drug Targeting; FDA approved; Foundations; Frequencies; Genes; Genetic Transcription; Goals; Growth; Growth and Development function; Helicobacter pylori; Human; Immunoprecipitation; Knowledge; Life Cycle Stages; Mass Spectrum Analysis; Mission; Molecular; Molecular Machines; Nucleotides; Outcome; Patients; Peptic Ulcer; Pharmaceutical Preparations; Preventive; Proteins; Proteolysis; Public Health; Regulon; Replication Initiation; Replication Origin; Research; Risk Factors; Sampling; Site; Techniques; Testing; Therapeutic; Translations; United States National Institutes of Health; Work; antimicrobial drug; cdc Genes; cell growth; cellular development; chromatin immunoprecipitation; chromosome replication; crosslink; deep sequencing; event cycle; in vivo; innovation; malignant stomach neoplasm; new therapeutic target; novel; pathogen; promoter; protein complex; tool; transcription factor

Project Summary/Abstract There is a fundamental gap in our understanding of how helicobacter pylori (Hp), a bacterium that is highly relevant to public health, regulates its life cycle. Although mechanistic details of how Hp induces damage to host cells have been revealed, our understanding for how this pathogen regulates its own proliferation remains unclear. The existence of this knowledge gap represents a burden in the need to develop novel specie-specific drugs that can control the proliferation of Hp in human patients. Our long-term goal is to identify drug targets that are essential in regulating Hp cell cycle and proliferation. The overall objective of the proposed research is to define the regulatory circuitry that controls the forward progression of Hp cell cycle. Our central hypothesis is that the chromosome replication initiator DnaA in Hp is a central node connecting multiple cell cycle regulating factors. DnaA is a multifunctional protein that aside from opening the chromosomal region known as the origin of replication (ori), it also acts as a highly regulated transcription factor. The rationale for the proposed research is that DNA replication initiators have been shown to influence the activity of other cellular machines involved in the cell cycle, such as cytokinesis, cell growth, and cellular development. Our plan is to test our central hypothesis, and thus accomplish our overall objective for this project, by pursuing the following three specific aims: 1) Define the DnaA transcriptional regulon by analyzing the global chromosome profile of promoters bound by DnaA using chromatin immunoprecipitation assays followed by deep sequencing; 2) Identify regulators that modulate DnaA's activity at the origin of replication by cross-linking and isolating the chromosomal ori-proteins complex using a DNA-sampling technique; 3) Identify factors that directly interact with DnaA by isolating DnaA-containing protein complexes within the cell. Successful completion of the proposed research is expected to vertically advance and expand our knowledge of and ability to assemble the regulatory circuitry that controls Hp cell cycle. Furthermore, findings from our work will broaden our understanding of how the highly conserved replication initiators coordinate DNA synthesis with the progression of the cell cycle. Our proposed research is innovative, in our opinion, because it represents a substantive departure from the status quo by targeting Hp-specific cell cycle regulators as targets for new antibiotics.

项目摘要/摘要 我们对幽门螺杆菌（HP）的理解有一个根本的差距，这是一种高度的细菌 与公共卫生有关，调节其生命周期。尽管HP如何造成损害的机械细节 已经揭示了宿主细胞，我们对这种病原体如何调节其自身增殖的理解 不清楚。这种知识差距的存在代表了需要开发新颖特定特定规格的负担 可以控制人类患者HP增殖的药物。我们的长期目标是确定药物目标 这对于调节HP细胞周期和增殖至关重要。拟议研究的总体目的是 定义控制HP细胞周期的正向进程的调节电路。我们的中心假设是 HP中的染色体复制引发剂DNAA是连接多个细胞周期调节的中央节点 因素。 DNAA是一种多功能蛋白，除了打开称为起源的染色体区域 在复制（ORI）中，它也充当高度调节的转录因子。提议的理由 研究表明，DNA复制启动器已被证明会影响其他细胞机器的活性 参与细胞周期，例如细胞因子，细胞生长和细胞发育。我们的计划是测试我们的 中心假设，从而实现我们对该项目的整体目标，通过追求以下三个 具体目的：1）通过分析全局染色体曲线来定义DNAA转录调节 使用染色质免疫沉淀测定法结合的启动子，然后进行深层测序。 2） 通过交联和隔离来确定调节DNAA活性在复制起源的调节剂 使用DNA采样技术的染色体原蛋白复合物； 3）确定直接相互作用的因素 与DNAA通过分离细胞内的含DNAA的蛋白质复合物。成功完成 预计拟议的研究将垂直提高和扩大我们对组装的知识和能力 控制HP细胞周期的调节电路。此外，我们的工作发现将扩大我们的 了解高度保守的复制启动器如何与进展协调DNA合成 细胞周期。在我们看来，我们提出的研究具有创新性，因为它代表了实质性 通过将HP特异性细胞周期调节剂作为新抗生素的靶标，与现状不同。