Targetomes of infection-specific small RNAs of Bartonella bacilliformis

巴尔通氏杆菌(Bartonella bacilliformis，BB)是一种高毒力的沙蝇媒介细菌，可引起卡里翁病。 人类。这种长期被忽视的热带疾病正在安第斯山脉和 扩展到秘鲁、厄瓜多尔和墨西哥的非流行区(即低海拔和更多样化的生境) 哥伦比亚。据估计，在南美洲约14.5万平方公里的区域内，有200万人面临着 在疫区报告的脑炎发病率为12.7/100人年。bb 感染具有潜在的生命威胁，如果不治疗，死亡率高达88%，约10%的死亡率 抗菌干预后的发生率。在非流行地区，卡里翁病表现为急性 发热性疾病，红细胞压积下降约80%(奥罗亚热)，而在流行地区， 血管瘤性皮肤损害(Peruana)和慢性菌血症是最常见的，有效地造成了 人类是病原体的蓄水池。我们的长期目标是阐明BB是如何调节基因表达的 在沙蝇媒介和人类宿主中生存所需的产品，总体目标为：a) 产生可应用于最终根除卡里翁病的信息，以及b)提供 巴尔通体和其他节肢动物传播的致病细菌的毒力基因调控模型。我们的 发现24个人类感染特异性sRNA和8个沙蝇特异性sRNA(总共160个BB sRNA 已鉴定)表明sRNA用于转录后控制宿主和载体中的翻译， 分别进行了分析。我们假设人类感染特异的反式作用的sRNA调节基因的转录本。 人类脑瘫生存所需的产品。这一假设将通过两个具体目标来解决。在……里面 目的1，我们将通过对三个重要的sRNA的交联-序列分析来鉴定它们的mRNAs靶体。 BB现场直播。在目标2中，我们将使用EMSA和报告程序来确认和分析已识别的sRNA-mrna相互作用 构造。这项研究的结果将为三种主要的感染规律提供基础分析-- Bb的特异性sRNA。更广泛地说，这项研究还将建立一个模型，以阐明sRNA调节基因在 节肢动物携带的细菌病原体。

Bartonella bacilliformis (Bb) is a highly-virulent, sand fly-vectored bacterium responsible for Carrión’s disease in humans. This chronically-neglected tropical disease is re-emerging in endemic regions of the Andes and expanding into non-endemic areas (i.e., lower altitudes and more diverse habitats) of Peru, Ecuador and Colombia. An estimated two million people in an area of ~145,000 square-km in South America are at risk of infection by Bb, and incidence rates of 12.7 / 100 person-years have been reported in endemic regions. Bb infections are potentially life-threatening, with fatality rates as high as 88% if left untreated, and ~10% fatality rates following antimicrobial intervention. In non-endemic regions, Carrión’s disease manifests as an acute febrile illness with an ~80% reduction in erythrocyte hematocrit (Oroya fever), whereas in endemic regions, angiomatous skin lesions (verruga peruana) and chronic bacteremia are most common, effectively creating a human reservoir for the pathogen. Our long-term goal is to elucidate how Bb regulates expression of gene products required to survive in the sand fly vector and human host, with overarching objectives of: a) generating information that can be applied to the eventual eradication of Carrión's disease and b) providing a model of virulence gene regulation in Bartonella and other arthropod-vectored, pathogenic bacteria. Our discovery of 24 human-infection-specific sRNAs and 8 sand fly-specific sRNAs (from a total of 160 Bb sRNAs identified) suggests that sRNAs are used to post-transcriptionally control translation in the host and vector, respectively. We hypothesize that human-infection-specific, trans-acting sRNAs regulate transcripts for gene products that are required for Bb survival in humans. The hypothesis will be addressed by two specific aims. In aim 1, we will characterize the mRNA targetomes of three prominent sRNAs through crosslink-Seq analysis of live Bb. In aim 2, we will confirm and analyze the identified sRNA-mRNA interactions using EMSAs and reporter constructs. Results of the study will provide a foundational analysis of the regulons for three dominant, infection- specific sRNAs of Bb. More broadly, the research will also establish a model for elucidating sRNA regulons in arthropod-vectored bacterial pathogens.