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.

项目总结 莱姆病的病原体伯氏疏螺旋体是由节肢动物传播的 美国每年有超过47万例确诊病例。鉴于莱姆病是一个重要的公众 健康问题，研究BB如何建立和维持感染对于评估其 致病潜力和制定预防或治疗策略。众所周知，BB修改了它的 在其致病生命周期中，特别是在节肢动物传播期间的转录图谱 传播到哺乳动物身上。BosR是一种重要的转录调控因子，其功能类似于全球金属调节因子 蛋白质，并改变哺乳动物定居所需基因的表达。具体地说，需要BosR 对于小鼠的实验性感染，是激活rpos/RpoN/RRP2级联反应所必需的， 基因无关基因的表达，包括BB感染所需的毒力决定因素。而当 毫无疑问，依赖于BosR的rpos调控对疏螺旋体的传染性很重要，有 BosR直接或间接调节的其他基因可能是BB感染的因素。BosR是怎样的 与这一复杂的监管网络相关联的问题尚未得到彻底解决。在这里，几部小说博斯尔 描述了迄今为止未知的调节功能，包括BosR对RNA的识别。这个 初步数据显示，BosR与小分子非编码RNA结合，并显示出伴侣活性。 BosR与sRNA形成一个复合体，它与BB转录本结合，并针对它们进行降解或增强 翻译。BosR的这种转录后调控形式代表了一种新的创新的调控 该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，鉴定了BosR：：SRNA复合体识别的转录本，并与BosR相关的 SRNAs对斑疹病毒的致病作用已确立。在目标2中，将挖掘最新的CHIP-SEQ数据来评估新的BosR DNA结合。初步结合研究证实了两个目标的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的结合。将分析BosR DNA结合以 确定BosR的氧化还原和金属结合状态如何改变识别的目标序列。独一无二 将跟踪BosR结合基序，并确定一致的序列。最后，实验将是 用来确定异位产生rpos的BosR突变体的表型以质疑直接作用 脑脊液感染中的BosR。综上所述，这项提案将为BOR如何调解提供新的理解 转录和转录后调控随之而来，并对BosR如何整合氧化还原提供了新的见解 BB的状态影响核酸结合和斑疹病毒的发病。