A small RNA regulation of virulence in Lyme disease pathogen

莱姆病病原体毒力的小RNA调节

• 批准号: 10218725
• 负责人: Xiuli Yang
• 金额: $ 7.73万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10218725
• 关键词: Address; Affect; Antibiotics; Arbovirus Infections; Area; Bacteria; Bacteria sigma factor KatF protein; Base Pairing; Binding; Binding Proteins; Biological; Biological Process; Biology; Body Temperature; Borrelia; Borrelia burgdorferi; Cells; Complex; Data; Development; Environment; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genetic Translation; Genome; Goals; Impairment; Infection; Intercistronic Region; Intergenic DNA; Ixodes; Kanamycin; Kanamycin A; Life Cycle Stages; Location; Lyme Disease; Mammals; Measures; Messenger RNA; Microbe; Microbial Physiology; Mus; Mutagenesis; Nature; Northern Blotting; Nucleotides; Open Reading Frames; Order Spirochaetales; Organism; Pathogenesis; Play; Preventive; Proteins; RNA; RNA Binding; RNA-Binding Proteins; Regulation; Regulator Genes; Regulon; Reporting; Reverse Transcriptase Polymerase Chain Reaction; Role; Seminal; Series; Small RNA; Stimulus; Temperature; Therapeutic; Ticks; Tissues; Untranslated RNA; Untranslated Regions; Virulence; Work; base; differential expression; environmental change; enzootic; mRNA Stability; microbial; pathogen; pathogenic bacteria; response; tissue tropism; transmission process

Project Summary/Abstract Lyme disease, a prevalent arthropod-borne disease, is caused by the bacterial pathogen Borrelia burgdorferi. The microbes live in an intricate enzootic life cycle involving both Ixodes ticks and diverse mammalian species. To survive in its complex cycle and navigate through disparate sets of tissue environments, such as ones in mammalian hosts or in ticks, B. burgdorferi must regulate gene expression in a tight temporal and spatial manner, although the mechanism of such gene regulation remains unclear. Posttranscriptional gene regulation via a variety of small regulatory RNAs (sRNAs) is one mechanism of modulating gene expression in bacteria. Base pairing between the sRNAs and target mRNAs can trigger alterations in mRNA translation and stability, thereby influencing target gene expression. The small RNAs also bind to proteins, either sequestering or affecting the activities of proteins. Previous studies have reported that a small RNA, DsrABb, regulates RpoS gene expression in B. burgdorferi. Moreover, recent studies identified over 1000 B. burgdorferi sRNAs that are differentially regulated by environmental conditions, suggesting that sRNAs may play a role in gene regulation for spirochete adaptation during its enzootic cycle, although their precise functions or importance in spirochete biology and infectivity remain largely enigmatic. Most recently, an intergenic non-coding small RNA located upstream of bbd18, called ittA, was characterized and shown to be required for optimal infectivity and tissue tropism in B. burgdorferi. In fact, the intergenic region of bbd17 and bbd18 was reported to harbor a small RNA 0735 (SR0735), that is highly expressed at 37°C (mammalian host body temperature). Our preliminary data showed that the insertion of a kanamycin cassette upstream of SR0735 affects B. burgdorferi infectivity and decreases the level of SR0735, as shown in Northern blot and RT-PCR analyses, suggesting a critical importance of SR0735 in microbial virulence. The goals of our current proposal are to study the potential role of this sRNA in spirochete biology and virulence, and to identify its potential target RNA and/or protein binding partners. This study will uncover the sRNA regulatory mechanisms in spirochete biology and may help to decipher the functions of sRNAs in spirochete virulence and pathogenesis.

项目摘要/摘要 莱姆病是一种流行的节肢动物传播疾病，由伯氏疏螺旋体细菌引起。 这些微生物生活在一个错综复杂的地方性生活史中，包括硬蜱和不同的哺乳动物物种。 要在复杂的循环中生存并在不同的组织环境中导航，例如在 无论是在哺乳动物宿主还是在硬蜱体内，伯氏杆菌都必须在严格的时间和空间上调节基因的表达。 虽然这种基因调控的机制尚不清楚。转录后基因 通过多种小分子调控RNA(SRNAs)调控基因表达是一种重要的调控机制 细菌。SRNAs和靶mRNAs之间的碱基配对可以触发mRNAs翻译和 稳定性，从而影响靶基因的表达。小核糖核酸也能与蛋白质结合，要么隔离 或影响蛋白质的活性。以前的研究已经报道了一个小的RNA，DsrABb，调节rpos 伯氏杆菌的基因表达。此外，最近的研究发现，超过1000个伯氏杆菌sRNA是 受环境条件的差异调控，提示sRNA可能在基因调控中发挥作用 对于螺旋体在其地方性循环中的适应，尽管它们在螺旋体中的确切功能或重要性 生物学和传染性在很大程度上仍然是个谜。最近，一个基因间的非编码小RNA被定位 Bbd18的上游，称为itta，其特征表明是获得最佳传染性和组织所必需的。 伯氏假单胞菌的嗜性。事实上，据报道，bbd17和bbd18的基因间隔区含有一个小的 RNA 0735(SR0735)，在37℃(哺乳动物宿主温度)下高表达。我们的预赛 数据显示，在SR0735上游插入卡那霉素盒会影响伯氏杆菌的感染性 并降低SR0735的水平，Northern印迹和RT-PCR分析显示，这表明关键的 SR0735在微生物毒力中的重要性。我们当前提案的目标是研究潜在的作用 确定该sRNA在螺旋体生物学和毒力中的作用，并确定其潜在的靶向RNA和/或蛋白质结合。 合伙人。这项研究将揭示螺旋体生物学中的sRNA调节机制，并可能有助于 破译sRNA在螺旋体毒力和致病机制中的功能。