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)的理解存在一个根本性的空白，幽门螺杆菌是一种高度敏感的细菌 与公共健康相关，规范其生命周期。尽管惠普如何导致损害的机械细节 宿主细胞已经被揭示，我们对这种病原体如何调节自身增殖的了解仍然存在 不清楚。这种知识鸿沟的存在代表了开发新物种专属的需要的负担。 可以控制人类患者幽门螺杆菌增殖的药物。我们的长期目标是确定药物靶标 在调节幽门螺杆菌细胞周期和增殖过程中起着至关重要的作用。拟议研究的总体目标是 确定控制幽门螺杆菌细胞周期正向进展的调节电路。我们的中心假设是 幽门螺杆菌染色体复制启动子DNAA是连接多种细胞周期调控的中心节点 各种因素。DNAA是一种多功能蛋白质，除了打开被称为起源的染色体区域外， 复制因子(ORI)，它也是一个高度受调控的转录因子。建议的理由是 研究表明，DNA复制启动子可以影响其他细胞机器的活动 参与细胞周期，如胞质分裂、细胞生长和细胞发育。我们的计划是测试我们的 中心假设，从而实现我们这个项目的总体目标，实现以下三个目标 具体目标：1)通过分析全球染色体图谱，确定Dna A转录调节子 用染色质免疫沉淀分析DNAA结合的启动子，然后进行深度测序； 确定在复制起始处通过交联和分离DNAA的活性来调节DNAA活性的调节子 使用DNA采样技术的染色体或蛋白质复合体；3)确定直接相互作用的因素 通过分离细胞内含有DNAA的蛋白质复合体与DNAA结合。圆满完成 拟议的研究有望垂直推进和扩大我们对组装 控制HP细胞周期的调节电路。此外，我们的工作成果将扩大我们的 理解高度保守的复制启动子如何协调DNA合成与进程 对细胞周期的影响。在我们看来，我们提出的研究是创新的，因为它代表了 改变现状，将幽门螺杆菌特有的细胞周期调节因子作为新抗生素的靶标。