Post-translational regulation of alpha PSM production in Staphylococcus aureus by the small RNA Teg41

小 RNA Teg41 对金黄色葡萄球菌中 α PSM 产生的翻译后调节

• 批准号: 10461882
• 负责人: Ronan Carroll
• 金额: $ 41.04万
• 依托单位: OHIO UNIVERSITY ATHENS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10461882
• 关键词: Abscess; Amino Acids; Attention; Base Pairing; Binding; Biochemical; Biochemical Genetics; Biological; Biological Assay; Cell Wall; Cells; Classification; Communities; Dangerousness; Data; Disease; Genetic; Genetic Transcription; Health; High-Throughput DNA Sequencing; Human; Human body; In Vitro; Infection; Mediating; Messenger RNA; Modeling; Molecular; Molecular Biology; Molecular Conformation; Mus; Nucleotides; Operon; Pathogenesis; Pathogenicity; Peptides; Phenols; Play; Post-Transcriptional Regulation; Post-Translational Regulation; Production; Proteomics; RNA; Regulation; Role; Skin Tissue; Small RNA; Soft Tissue Infections; Staphylococcus aureus; Staphylococcus aureus infection; Structure; Suggestion; Techniques; Testing; Therapeutic Intervention; Toxin; Transcript; Translations; United States; Variant; Virulence; Virulence Factors; Work; attenuation; base; defined contribution; experimental study; in silico; in vivo; methicillin resistant Staphylococcus aureus; new therapeutic target; pathogen; pathogenic bacteria; ribosome profiling; virtual

PROJECT SUMMARY/ABSTRACT Staphylococcus aureus is both a commensal of humans and a highly dangerous bacterial pathogen. S. aureus pathogenesis is mediated by a large repertoire of secreted and cell wall-associated virulence factors, including a number of potent cytolytic peptides called phenol soluble modulins (PSMs). PSMs are amphipathic, alpha helical peptides that vary in size depending on their classification. The α type PSMs are ~22 amino acids in size and have been the focus of intense study in recent years. They have been implicated in contributing to the high virulence potential of community-acquired methicillin resistant S. aureus (CA-MRSA) strains, in particular those of the USA300 lineage. While the role of αPSMs in S. aureus infection has been extensively investigated, significant gaps still exist in our understanding of how they are produced in the bacterial cell. There are five αPSM peptides produced by most S. aureus strains (PSMα1-4 and the δ-toxin). PSMα1-4 are encoded within the same polycistronic transcript (the αPSM transcript), yet studies have shown that the relative levels of the four peptides vary considerably. PSMα4 (located at the 3' end of the transcript) is commonly the most abundant, while PSMα3 (the most potent of the four peptides) is typically the least abundant. This variation in αPSM abundance is suggestive of post-transcriptional regulation. The long-term objective of this project is to understand the molecular mechanism(s) that contribute to αPSM production in S. aureus. In this proposal, we will specifically investigate the contribution of the small RNA Teg41 to αPSM production and virulence. Preliminary studies show that Teg41 positively influences αPSM production at the post-transcription level. We will investigate (i) which of the PSMα1-4 peptides is/are regulated by Teg41, (ii) if Teg41-mediated regulation is facilitated by direct base pairing with the αPSM transcript, and (iii) at what stage in αPSM production, and how, Teg41 exerts its influence. To investigate these three aims, we will use a combination of in vitro, in vivo, genetic, biochemical, and molecular biology approaches. We will also utilize a number of cutting edge techniques based on high throughput DNA sequencing (SHAPE-seq, miR-CATCH, and Ribo-seq). The results from this study could have direct implications for human health. Disrupting Teg41-mediated αPSM-production could dramatically lower the virulence potential of S. aureus and therefore may represent a novel target for therapeutic intervention.

项目概要/摘要 金黄色葡萄球菌既是人类的共生菌，也是一种高度危险的细菌病原体。金黄色葡萄球菌 发病机制是由大量分泌的和细胞壁相关的毒力因子介导的，包括 许多有效的溶细胞肽，称为苯酚可溶性调节蛋白（PSM）。 PSM 是两亲性的，α 螺旋肽的大小根据其分类而变化。 α 型 PSM 的大小约为 22 个氨基酸 并成为近年来研究的热点。他们涉嫌为高额捐款做出贡献 社区获得性耐甲氧西林金黄色葡萄球菌 (CA-MRSA) 菌株的毒力潜力，特别是那些 USA300 血统。虽然 αPSM 在金黄色葡萄球菌感染中的作用已被广泛研究， 我们对它们如何在细菌细胞中产生的理解仍然存在重大差距。有五个 大多数金黄色葡萄球菌菌株产生的 αPSM 肽（PSMα1-4 和 δ-毒素）。 PSMα1-4 编码在 相同的多顺反子转录本（αPSM转录本），但研究表明这四种转录本的相对水平 肽差异很大。 PSMα4（位于转录本的 3' 端）通常是最丰富的， 而 PSMα3（四种肽中最有效的）通常是含量最少的。 αPSM 的这种变化 丰度表明存在转录后调控。该项目的长期目标是 了解金黄色葡萄球菌中促进 αPSM 产生的分子机制。在这个提案中，我们 将专门研究小RNA Teg41对αPSM产生和毒力的贡献。 初步研究表明，Teg41 在转录后水平对 αPSM 的产生产生积极影响。我们 将研究 (i) 哪些 PSMα1-4 肽受 Teg41 调节，(ii) Teg41 介导的调节是否受 Teg41 调节 通过与 αPSM 转录物直接碱基配对来促进，以及 (iii) 在 αPSM 生产的哪个阶段以及如何， Teg41发挥其影响力。为了研究这三个目标，我们将结合使用体外、体内、遗传、 生物化学和分子生物学方法。我们还将利用一些基于的尖端技术 高通量 DNA 测序（SHAPE-seq、miR-CATCH 和 Ribo-seq）。这项研究的结果可以 对人类健康有直接影响。破坏 Teg41 介导的 αPSM 生产可能会显着降低 金黄色葡萄球菌的毒力潜力，因此可能代表治疗干预的新目标。