cAMP signaling in Mycobacterium tuberculosis

结核分枝杆菌中的 cAMP 信号传导

• 批准号: 7380040
• 负责人: Kathleen A McDonough
• 金额: $ 27.76万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-15 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7380040
• 关键词: Address; Adenylate Cyclase; Bacteria; Binding; Biological Assay; Condition; Cyclic AMP; DNA Microarray Chip; DNA Microarray format; Defect; Diagnostic; Disease; Environment; Foundations; Future; Gene Expression; Gene Expression Regulation; Genes; Hypoxia; Infection; Knock-out; Lead; Measures; Mediating; Molecular Genetics; Mycobacterium tuberculosis; Promoter Regions; Purpose; Range; Regulation; Regulator Genes; Regulon; Relative (related person); Reporter; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Signal Transduction; Therapeutic; Tuberculosis; Tuberculosis Vaccines; Virulence; Work; interdisciplinary approach; macrophage; member; mutant; prevent; programs; promoter; response; trafficking; transcription factor

DESCRIPTION (provided by applicant): The long term objective of this project is to better understand how Mycobacterium tuberculosis (Mtb) establishes infection so that effective strategies can be developed to prevent it. Gene regulation is a critical aspect of Mtb's successful response to the host environment during infection. The recent finding that Mtb encodes as many as 15 adenylate cyclases (AC) suggests that cAMP signaling constitutes an important, but previously unrecognized, regulatory mechanism in Mtb. A multidisciplinary approach will address the role of cAMP signaling in Mtb, at the molecular, genetic and cellular levels, with a focus on conditions associated with latency and Mtb's interaction with macrophages. Specific aims include: Aim 1: identify, using DMA microarrays and isogenic knockout mutants, Mtb genes that are regulated by the putative cAMP-dependent transcription factor, Rv1675c, at various levels of cAMP during hypoxia; Aim 2: define and characterize a second likely cAMP-dependent regulon controlled by putative transcription factor Rv3676 in Mtb. Candidate members of this regulon will be functionally assessed by defining the role of Rv3676 binding to a DMA motif identified in their promoter regions; Aim 3: evaluate the roles of cAMP and a mammalian-type AC, Rv1625c, in Mtb's interaction with macrophages, particularly with respect to bacterial replication and trafficking; Aim 4: assess the regulation and function of a specific macrophage-induced, cAMP regulated gene within macrophages and during latency-associated conditions. Identification and characterization of cAMP-mediated gene regulation in Mtb will contribute to our understanding of the factors needed for the establishment of tuberculosis infection and disease. This work will explore a new gene regulatory network in Mtb and provide an important foundation for future work on the role of cAMP signaling in Mtb virulence gene regulation, leading to identification of potential targets for TB vaccines, therapeutics, and/ or diagnostic purposes.

描述（由申请人提供）：该项目的长期目标是更好地了解结核分枝杆菌（MTB）如何建立感染，以便可以制定有效的策略来防止其。基因调节是MTB在感染过程中成功反应宿主环境的关键方面。最近的发现，MTB编码多达15个腺苷酸环化酶（AC）表明，cAMP信号传导构成了MTB中的重要但未被认可的调节机制。多学科方法将解决cAMP信号在MTB，分子，遗传和细胞水平上的作用，重点是与潜伏期和MTB与巨噬细胞的相互作用相关的条件。具体目的包括： AIM 1：使用DMA微阵列和同基因敲除突变体，MTB基因受推定的cAMP依赖性转录因子RV1675C调节的MTB基因，在缺氧期间的各个cAMP上。 AIM 2：定义并表征由MTB中推定的转录因子RV3676控制的第二个可能的CAMP依赖性调节。该法规的候选成员将通过定义RV3676与其启动子区域中鉴定的DMA基序的作用来评估功能。 AIM 3：评估CAMP和哺乳动物型AC RV1625C在MTB与巨噬细胞的相互作用中的作用，尤其是在细菌复制和运输方面； AIM 4：评估特定巨噬细胞诱导的cAMP调节基因在巨噬细胞和潜伏期相关条件下的调节和功能。 MTB中营地介导的基因调节的鉴定和表征将有助于我们理解建立结核病感染和疾病所需的因素。这项工作将探索MTB中的一个新基因调节网络，并为cAMP信号在MTB毒力基因调节中的作用提供了重要的基础，从而鉴定了针对TB疫苗，治疗剂和/或诊断目的的潜在靶标。