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: 10240573
• 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/10240573
• 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.

项目摘要/摘要 金黄色葡萄球菌既是人类的共生，又是高度危险的细菌病原体。 S.金黄色葡萄酒 发病机理是由大量分泌和细胞壁相关病毒因素的大量曲目介导的，包括 许多称为苯酚固体模量（PSM）的潜在胞溶肽。 PSM是两亲性的，Alpha 螺旋辣椒的大小根据其分类而变化。 α型PSM的大小约为22个氨基酸 近年来一直是激进研究的重点。他们被暗示是为了贡献高 社区获得的甲氧西林抗性金黄色葡萄球菌（CA-MRSA）菌株的病毒潜力，特别是 USA300血统。尽管已经广泛研究了αPSM在金黄色葡萄球菌感染中的作用，但 在我们对细菌细胞中它们如何产生的理解中，仍然存在很大的差距。有五个 大多数金黄色葡萄球菌菌株（PSMα1-4和δ-毒素）产生的αPSM肽。 PSMα1-4在 相同的多元转录本（αPSM转录本），但研究表明，四个相对水平 肽谨慎不同。 PSMα4（位于转录本的3'末端）通常是最丰富的， 虽然PSMα3（四个胡椒体中最潜力）通常是最少的。 αPSM的这种变化 抽象暗示了转录后调节。该项目的长期目标是 了解有助于金黄色葡萄球菌产生αPSM的分子机制。在这个建议中，我们 将特别研究小的RNA TEG41对αPSM产生和病毒的贡献。 初步研究表明，TEG41在转录后水平上积极影响αPSM的产生。我们 将研究（i）哪种PSMα1-4肽是由TEG41调节的，（ii）如果TEG41介导的调节为 通过直接碱基与αPSM转录物配对，以及（iii）在αPSM产生的哪个阶段以及如何促进，以及如何 TEG41发挥其影响力。为了研究这三个目标，我们将使用体外，体内，遗传， 生化和分子生物学方法。我们还将利用许多基于尖端的技术 在高吞吐量DNA测序（Shape-seq，mir-Catch和Ribo-Seq）上。这项研究的结果可能 对人类健康具有直接影响。破坏TEG41介导的αPSM产生可能会大大降低 金黄色葡萄球菌的病毒潜力可能代表治疗干预的新靶标。