SpoVG and PlzA Regulation of Lyme Disease Spirochete Infection Processes

SpoVG 和 PlzA 对莱姆病螺旋体感染过程的调节

• 批准号: 10597629
• 负责人: Brian Stevenson
• 金额: $ 69.78万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10597629
• 关键词: Affect; Affinity; Anti-Bacterial Agents; Antibiotic Therapy; Antibiotics; Antigenic Variation; Back; Bacteria; Bacterial Infections; Bacterial Physiology; Binding; Binding Proteins; Binding Sites; Biology; Borrelia; Borrelia burgdorferi; Cessation of life; ChIP-seq; Collaborations; Cues; DNA; DNA Binding; Data; Development; Environment; Failure; Gene Expression; Genes; Genetic Transcription; Genome; Homologous Gene; Human; Immunocompetent; Impairment; Infection; Infection Control; Infectious Agent; Invaded; Investigation; Joints; Knowledge; Laboratory Finding; Lyme Disease; Mammals; Maps; Mechanics; Membrane Proteins; Mus; Nature; North America; Order Spirochaetales; Organism; Pathogenesis; Pathogenicity; Patients; Periodicity; Physiology; Preventive therapy; Process; Production; Property; Proteins; Proteomics; RNA; RNA Sequences; RNA immunoprecipitation sequencing; RNA-Binding Proteins; Recording of previous events; Regulation; Regulatory Pathway; Regulon; Repression; Research Personnel; Role; Site; Specificity; Spirochaetales Infections; Ticks; Vertebrates; Virulence; Virulence Factors; Work; arthropod-borne; curative treatments; genetic regulatory protein; human disease; human pathogen; improved; insight; member; microbial; mutant; novel therapeutics; nucleic acid binding protein; pathogen; pathogenic bacteria; posttranscriptional; protein expression; response; stem; tick feeding; tick transmission; transcriptome; transcriptomics; transmission process; vector-borne pathogen

v ABSTRACT Bacterial infections require that the pathogen accurately produce essential factors at appropriate levels during each stage of infection processes. Understanding how bacteria control levels of their proteins in response to cues from their hosts provides important insights on microbial infectious properties. Such knowledge can also reveal new targets for improved preventative and curative therapies. The Lyme disease spirochete, Borrelia burgdorferi, survives in nature through cycles of infecting vertebrates and ticks. The CDC calculates that there are approximately 300,000 new cases of human Lyme disease in the USA each year. B. burgdorferi can persistently infect immunocompetent humans and other mammals for many years. Failure to treat Lyme disease promptly and adequately can result in persistent debilitating effects or, sometimes, death. Long-term infections may require extensive periods of antibiotic treatment. In order to better treat Lyme disease, it is critical to develop a more thorough understanding of B. burgdorferi biology, including the mechanisms by which the spirochete controls production of virulence factors. We discovered that a borrelial protein, SpoVG, binds with specificity and high affinity to DNA and RNA. Deletion of spoVG significantly impaired B. burgdorferi's ability to colonize ticks and be transmitted from ticks to mammals. Dysregulation of spoVG transcription caused significant changes in bacterial physiology. We further found that SpoVG directly interacts with another B. burgdorferi protein, PlzA, the Lyme spirochete's cyclic-di-GMP-binding protein. Our studies revealed that PlzA is also a site-specific nucleic acid-binding protein, and ΔplzA mutants are defective in their infectivity. Among the many regulated targets we identified for SpoVG and PlzA is the antigenically-variable VlsE surface protein, which is essential for persistent B. burgdorferi infection. The planned studies will simultaneously investigate SpoVG and PlzA, as well as the effects of c-di-GMP on their functions. These preliminary data stemmed from long-standing collaborations between the P.I. and co- investigators of this proposal. Our combined efforts will yield a comprehensive view of the mechanisms through which B. burgdorferi controls production of these critical regulatory factors, and deep insights on how SpoVG, PlzA, and c-di-GMP regulate VlsE and other virulence-associated proteins. In addition, many other pathogenic bacteria produce homologs of SpoVG, and our observations on B. burgdorferi SpoVG bear similarities with the protein's known effects in other bacterial pathogens. To date, little is known about how SpoVG affects virulence and physiology in any bacterial species. The interactions we discovered between SpoVG and PlzA raise the possibilities that other bacteria also control SpoVG function through c-di-GMP-binding proteins. Results of the planned investigations will provide useful insights on the regulatory mechanisms and infectious properties of numerous important human pathogens.

v❖ 摘要 细菌感染需要病原体在感染过程中准确地产生适当水平的必需因子。 感染过程的每个阶段。了解细菌如何控制其蛋白质水平以应对 来自宿主的线索提供了关于微生物感染特性的重要见解。这样的知识还可以 揭示改进预防和治疗疗法的新目标。 莱姆病螺旋体伯氏疏螺旋体通过感染脊椎动物的周期在自然界中生存 和蜱虫。 CDC 计算出，美国大约有 30 万新发人类莱姆病病例 美国每年。伯氏疏螺旋体可以持续感染免疫功能正常的人类和其他哺乳动物许多 年。未能及时、充分地治疗莱姆病可能会导致持续的衰弱效应，或者， 有时，死亡。长期感染可能需要长时间的抗生素治疗。为了更好 治疗莱姆病，更深入地了解伯氏疏螺旋体生物学至关重要，包括 螺旋体控制毒力因子产生的机制。 我们发现疏螺旋体蛋白 SpoVG 与 DNA 和 RNA 具有特异性和高亲和力结合。 spoVG 的缺失显着损害了伯氏疏螺旋体定植蜱虫和从蜱虫传播的能力 对哺乳动物。 spoVG 转录失调引起细菌生理学的显着变化。我们 进一步发现 SpoVG 直接与另一种伯氏疏螺旋体蛋白 PlzA（莱姆螺旋体的蛋白）相互作用 环二 GMP 结合蛋白。我们的研究表明，PlzA 也是一种位点特异性核酸结合 蛋白和ΔplzA突变体的感染性存在缺陷。在我们确定的众多监管目标中 SpoVG 和 PlzA 是抗原可变的 VlsE 表面蛋白，对于持久性芽孢杆菌至关重要。 伯氏杆菌感染。 计划中的研究将同时研究 SpoVG 和 PlzA，以及 c-di-GMP 对 他们的职能。这些初步数据源于 P.I. 之间的长期合作。并共同 该提案的调查员。我们的共同努力将通过以下方式全面了解这些机制： 伯氏疏螺旋体控制这些关键调节因子的产生，并深入了解 SpoVG 如何 PlzA 和 c-di-GMP 调节 VlsE 和其他毒力相关蛋白。 此外，许多其他病原菌也产生 SpoVG 的同源物，我们对 B. 伯氏疏螺旋体 SpoVG 与该蛋白在其他细菌病原体中的已知作用有相似之处。迄今为止，很少 已知 SpoVG 如何影响任何细菌物种的毒力和生理学。我们的互动 SpoVG 和 PlzA 之间的发现提出了其他细菌也控制 SpoVG 功能的可能性 通过 c-di-GMP 结合蛋白。计划调查的结果将为以下方面提供有用的见解： 许多重要人类病原体的调节机制和感染特性。

项目成果

Quantitative analyses of interactions between SpoVG and RNA/DNA.

SpoVG 和 RNA/DNA 之间相互作用的定量分析。

• DOI: 10.1101/2023.02.06.527361
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Saylor,TimothyC;Savage,ChristinaR;Krusenstjerna,AndrewC;Jusufovic,Nerina;Zückert,WolframR;Brissette,CatherineA;Motaleb,Md;Schlax,PaulaJ;Stevenson,Brian
• 通讯作者: Stevenson,Brian

Gac Is a Transcriptional Repressor of the Lyme Disease Spirochete's OspC Virulence-Associated Surface Protein.

• DOI: 10.1128/jb.00440-22
• 发表时间: 2023-04-25
• 期刊: Journal of bacteriology
• 影响因子: 3.2