A novel mechanism of rifamycin resistance in Mycobacterium abscessus mediated by a putative helicase

由假定的解旋酶介导的脓肿分枝杆菌利福霉素耐药性的新机制

• 批准号: 10408177
• 负责人: Pallavi Ghosh
• 金额: $ 18.91万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-20 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10408177
• 关键词: ADP Ribose Transferases; Actinobacteria class; Acute respiratory failure; Affinity Chromatography; Antibiotic Resistance; Antibiotics; Bacillus subtilis; Bacteria; Bacterial RNA; Binding; Binding Proteins; Biological Assay; Chronic; Chronic Obstructive Pulmonary Disease; Clinical; Cystic Fibrosis; DNA; DNA-Directed RNA Polymerase; Dose; Elements; Enzymes; Exposure to; FDA approved; Future; Genes; Genetic Transcription; Genome; Genus Mycobacterium; Growth; Hypersensitivity; In Vitro; Infection; Investigation; Laboratories; Lead; Longitudinal Studies; Lung; Lung infections; Mediating; Microbial Antibiotic Resistance; Mutation; Mycobacterium Infections; Mycobacterium abscessus; Mycobacterium tuberculosis; Nightmare; Pathway interactions; Patients; Pharmaceutical Preparations; RNA chemical synthesis; Regulation; Replication Initiation; Reporter; Resistance; Rifabutin; Rifampicin resistance; Rifampin; Rifamycins; Skin Tissue; Soft Tissue Infections; Transcriptional Regulation; Tuberculosis; analog; antimicrobial; design; genetic regulatory protein; helicase; inhibitor; insight; lung injury; mouse model; mutant; non-tuberculosis mycobacteria; novel; prevent; resistance gene; resistance mechanism; therapeutic development; transcription factor

Project Summary: Mycobacterium abscessus (Mab) is a rapidly growing NTM causing skin and soft tissue infections as well as pulmonary infections in patients with chronic lung damage. Mab stands apart as one of the most antibiotic resistant microbial species, making its infections incredibly difficult to treat. Particularly striking is its resistance to rifampicin (RIF), a frontline drug for many mycobacterial infections including tuberculosis. RIF inhibits global RNA synthesis by binding to the b-subunit of bacterial RNA polymerase; additionally it is also known to inhibit replication initiation due to an inhibition of dnaA expression. The intrinsic RIF resistance in Mab has so far been attributed to the presence of an ADP-ribosyltransferase (Arr) activity that ribosylates RIF leading to its inactivation. However, we have recently identified an additional determinant, MAB_3189c - a putative helicase, that confers high levels of RIF resistance in Mab. MAB_3189c expression is RIF inducible and is likely regulated by a RIF-associated element (RAE) dependent transcription factor. In this project we will determine the regulation and function of Mab3189c in RIF resistance. In Aim 1 we will investigate if Mab3189c mediates RIF resistance by either directly protecting RNAP against the action of RIF thereby enabling global RNA synthesis to continue, or by alleviating RIF-sensitive replication initiation at oriM thereby enabling growth in the presence of RIF. In Aim 2 we propose to identify the regulatory protein required for RAE-dependent induction of MAB_3189c. The findings will provide a platform for a long-term study to gain insights into Mab3189c dependent RIF resistance in Mab, and design of new strategies to treat Mab infections.

项目摘要： 分枝杆菌（MAB）是一种快速生长的NTM，引起皮肤和软组织感染 以及慢性肺损伤患者的肺部感染。 mab是最多的人之一 抗生素耐药物种，使其感染难以治疗。特别是惊人的是 它对利福平（RIF）的耐药性，这是一种用于许多分枝杆菌感染的前线药物，包括结核病。 RIF通过与细菌RNA聚合酶的B-亚基结合来抑制全局RNA合成；另外 还已知由于DNAA表达抑制复制起始。内在的RIF 迄今为止，MAB中的抗性归因于ADP-核糖基转移酶（ARR）活性的存在 核糖基rif导致其失活。但是，我们最近确定了 决定符，mab_3189c-一种推定的解旋酶，它赋予了MAB中较高的RIF耐药性。 MAB_3189C的表达是rif诱导的，很可能由RIF相关元件（RAE）调节 依赖转录因子。在这个项目中，我们将确定MAB3189C的调节和功能 在RIF抗性中。在AIM 1中，我们将调查MAB3189C是否直接介导了RIF的耐药性 保护RNAP免受RIF的作用，从而使全球RNA合成继续或通过 减轻Orim的RIF敏感复制启动，从而在RIF的存在下实现增长。目标 2我们建议确定MAB_3189C RAE依赖性诱导所需的调节蛋白。这 调查结果将为长期研究提供一个平台，以洞悉MAB3189C依赖RIF MAB的耐药性，以及治疗MAB感染的新策略的设计。

项目成果

Mycobacterium abscessus HelR interacts with RNA polymerase to confer intrinsic rifamycin resistance.

• DOI: 10.1016/j.molcel.2022.06.034
• 发表时间: 2022-09-01
• 期刊: MOLECULAR CELL
• 影响因子: 16
• 作者: Hurst-Hess, Kelley R.;Saxena, Aavrati;Rudra, Paulami;Yang, Yong;Ghosh, Pallavi
• 通讯作者: Ghosh, Pallavi