Regulatory Pathways in Borrelial Pathogenesis

伯氏疏螺旋体发病机制的调节途径

• 批准号: 10677726
• 负责人: Jenny A. Hyde
• 金额: $ 66.31万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-05 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10677726
• 关键词: Address; Adhesions; Affect; Arbovirus Infections; Arthropod Vectors; Attenuated; Bacteria sigma factor KatF protein; Binding; Binding Sites; Biochemical; Borrelia; Borrelia burgdorferi; Borrelia oxidative stress regulator; ChIP-seq; Complement; Complex; Consensus Sequence; Cysteine; DNA; DNA Binding; DNA Sequence; Data; Diagnosis; Etiology; Event; Exhibits; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Homologous Gene; Infection; Infectious Agent; Life Cycle Stages; Link; Lyme Disease; Mammals; Mediating; Metals; Methodology; Molecular Chaperones; Mus; Nucleic Acid Binding; Oxidation-Reduction; Oxidative Stress; Pathogenesis; Pathogenicity; Pathway interactions; Persons; Phenotype; Post-Transcriptional Regulation; Prevention; Prevention strategy; Production; Promoter Regions; Proteins; Public Health; Publications; RNA; RNA Binding; Recording of previous events; Regulation; Regulator Genes; Regulatory Pathway; Regulon; Research; Role; Scheme; Structure; Testing; Therapeutic; Time; Transcript; Transcriptional Regulation; Translations; Uncertainty; United States; Untranslated RNA; Virulence; Work; arthropod-borne; crosslinking and immunoprecipitation sequencing; dimer; experience; experimental study; genetic regulatory protein; innovation; insight; interest; metalloregulatory protein; mutant; novel; skills; stem; tick transmission; transcriptome; transcriptome sequencing; transmission process; virulence gene

PROJECT SUMMARY The etiologic agent of Lyme Disease, Borrelia burgdorferi (Bb), is the leading arthropod-borne infection in the United States with over 470,000 cases diagnosed annually. Given that Lyme Disease is a significant public health concern, studies addressing how Bb establishes and maintains infection are vital to evaluating its pathogenic potential and developing prevention or therapeutic strategies. It is well known that Bb modifies its transcriptional profile throughout its pathogenic lifecycle, particularly during transmission from the arthropod vector into mammals. BosR is an important transcriptional regulator that functions as a global metalloregulatory protein and alters the expression of genes needed for mammalian colonization. Specifically, BosR is required for experimental infection in mice and is needed for the activation of the RpoS/RpoN/Rrp2 cascade that drives the expression of genetically unlinked genes, including virulence determinants required for Bb infection. While there is no doubt that the BosR-dependent regulation of rpoS is important to borrelial infectivity, there are additional genes that BosR regulates either directly or indirectly that may factor into Bb infection. How BosR is linked to this complex regulatory network has not been thoroughly addressed. Here, several novel BosR regulatory functions are described, heretofore unknown, including the recognition of RNA by BosR. The Preliminary Data shows that BosR binds to small non-coding RNA and exhibits chaperone activity suggesting that BosR forms a complex with sRNAs that binds to Bb transcripts and targets them for degradation or enhanced translation. This form of post-transcriptional regulation by BosR represents a new and innovative regulatory scheme that adds to its known DNA binding activity. In Aim 1, a subset of sRNAs that BosR binds will be evaluated, the transcripts recognized by the BosR::sRNA complex identified, and a link with the BosR-associated sRNAs to borrelial pathogenesis established. In Aim 2, recent ChIP-seq data will be mined to assess novel BosR DNA binding. Preliminary binding studies confirmed the ChIP-seq data for two targets and suggested that the redox status and metal coordination of BosR may alter DNA binding. BosR DNA binding will be analyzed to determine how the redox and metal binding status of BosR changes the target sequences recognized. Unique BosR binding motifs will be tracked, and a consensus sequence ascertained. Finally, experiments will be conducted to determine the phenotype of a bosR mutant that ectopically produces RpoS to query the direct role of BosR in Bb infection. Taken together, this proposal will provide new appreciation into how BosR-mediated transcriptional and post-transcriptional regulation ensues and novel insight into how BosR integrates the redox status of Bb to affect nucleic acid binding and borrelial pathogenesis.

项目概要 莱姆病的病原体伯氏疏螺旋体 (Bb) 是该地区主要的节肢动物传播的感染 美国每年确诊病例超过 47 万例。鉴于莱姆病是一种重要的公众疾病 由于健康问题，研究 Bb 如何建立和维持感染对于评估其感染至关重要 致病潜力并制定预防或治疗策略。众所周知，Bb修改了它的 整个致病生命周期的转录谱，特别是从节肢动物传播期间 载体进入哺乳动物。 BosR 是一种重要的转录调节因子，具有全局金属调节作用 蛋白质并改变哺乳动物定植所需基因的表达。具体来说，需要BosR 用于小鼠实验感染，并且是激活驱动 RpoS/RpoN/Rrp2 级联所必需的 遗传上不相关的基因的表达，包括 Bb 感染所需的毒力决定因素。尽管 毫无疑问，rpoS 的 BosR 依赖性调节对于疏螺旋体感染性很重要，有 BosR 直接或间接调节的其他基因可能会影响 Bb 感染。 BosR 怎么样 与这个复杂的监管网络相关的问题尚未得到彻底解决。这里，几个新颖的BosR 描述了迄今为止未知的调节功能，包括 BosR 对 RNA 的识别。这 初步数据显示 BosR 与小非编码 RNA 结合并表现出伴侣活性，表明 BosR 与 sRNA 形成复合物，与 Bb 转录物结合并靶向它们进行降解或增强 翻译。 BosR 的这种转录后监管形式代表了一种新的创新监管方式 方案增加了其已知的 DNA 结合活性。在目标 1 中，BosR 结合的 sRNA 子集将是 评估后，确定了 BosR::sRNA 复合物识别的转录本，以及与 BosR 相关的链接 sRNA 与疏螺旋体发病机制的关系已确立。在目标 2 中，将挖掘最近的 ChIP-seq 数据来评估新型 BosR DNA 结合。初步结合研究证实了两个靶标的 ChIP-seq 数据，并表明 BosR 的氧化还原状态和金属配位可能会改变 DNA 结合。 BosR DNA 结合将被分析 确定 BosR 的氧化还原和金属结合状态如何改变识别的靶序列。独特的 BosR 结合基序将被追踪，并确定共有序列。最后将进行实验 确定异位产生 RpoS 的 bosR 突变体的表型，以探究其直接作用 Bb 感染中的 BosR。总而言之，该提案将为 BosR 介导的方式提供新的认识 转录和转录后调控随之发生，并对 BosR 如何整合氧化还原有了新的见解 Bb 的状态影响核酸结合和疏螺旋体发病机制。