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.

项目总结/摘要 在我们对幽门螺杆菌（Hp）的理解方面存在根本性的差距，Hp是一种高度感染的细菌， 与公众健康相关，调节其生命周期。虽然Hp如何诱导损伤的机制细节， 尽管我们已经揭示了这种病原体是如何调节其自身增殖的， 不清楚这种知识差距的存在代表了开发新的物种特异性药物的需要的负担。 可以控制人类患者中Hp增殖的药物。我们的长期目标是确定药物靶点 在调节Hp细胞周期和增殖中至关重要。拟议研究的总体目标是 以确定控制Hp细胞周期向前进展的调节电路。我们的核心假设是 Hp染色体复制起始子DnaA是连接多个细胞周期调控的中心节点， 因素DnaA是一种多功能蛋白质，除了打开染色体上称为起点的区域外， 在复制（ori）中，它也作为高度调节的转录因子。建议的理由 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 中心假设，从而实现我们的总体目标，为这个项目，通过追求以下三个 具体目的：1）通过分析DNA的染色体分布， 使用染色质免疫沉淀测定，随后进行深度测序，由DnaA结合的启动子; 2） 通过交联和分离DnaA，鉴定在复制起点调节DnaA活性的调节剂。 染色体ori-蛋白质复合物，使用DNA取样技术; 3）鉴定直接相互作用的因子 通过分离细胞内含有DnaA的蛋白质复合物，成功完成 拟议的研究预计将垂直推进和扩大我们的知识和能力，组装 控制Hp细胞周期的调节电路。此外，我们工作的发现将扩大我们的 了解高度保守的复制启动子如何协调DNA合成， 细胞周期。我们认为，我们提出的研究是创新的，因为它代表了一个实质性的问题。 通过靶向Hp特异性细胞周期调节剂作为新抗生素的靶点来改变现状。