Investigating the extracellular RNA content of Staphylococcus aureus

项目总结/摘要 细胞外囊泡（EV）是由细胞产生和释放的纳米尺寸的膜封闭囊泡， 生活的领域。EV含有多种生物分子，例如脂质、蛋白质和核酸，并且是 取决于祖细胞，参与广泛的功能。革兰阳性菌EV的研究 细菌是一个相对较新的领域。由于革兰氏阳性细胞壁的结构， 革兰氏阳性菌不可能产生EV。然而，最近的研究表明，电动汽车生产在一个 各种革兰氏阳性菌和涉及EV的重要功能，如发病机制和免疫 逃避金黄色葡萄球菌既是人类的寄生虫，也是一种高度危险的细菌病原体。 生产S。金黄色葡萄球菌已经被证明，EV显示出细胞溶解性，并在免疫中发挥作用。 activation.研究S。金黄色葡萄球菌EV组合物集中在它们的蛋白质含量上，迄今为止， 报道描述了S.金黄色EV。S.已知金黄色葡萄球菌的DNA和RNA会引起 然而，这些核酸进入宿主细胞的机制是 未知我们假设EV代表了S.引入金黄色葡萄球菌DNA/RNA 进入宿主细胞在这个探索性的建议中，我们提出了初步的数据，表明RNA位于内部， 与S。金黄色EV。我们建议从五种不同的S. 金黄色葡萄球菌菌株，并使用该信息来定义S.金黄色葡萄球菌EV-RNome -即 在所有S.金黄色葡萄球菌菌株。定义EV RNome将允许我们识别候选 RNA分子用于未来的分析，以确定它们对EV免疫刺激特性的贡献。

PROJECT SUMMARY/ABSTRACT Extracellular vesicles (EVs) are nano sized, membrane-enclosed vesicles produced and released by cells in all domains of life. EVs contain a variety of biomolecules, such as lipids, proteins, and nucleic acids, and are involved in a broad range of functions depending on the progenitor cell. The study of EVs in Gram-positive bacteria is a relatively new field. Due to the architecture of the Gram-positive cell wall it was long thought to be impossible for Gram-positive bacteria to produce EVs. However recent studies have shown EV production in a variety of Gram-positive bacteria and implicated EVs in important functions like pathogenesis and immune evasion. Staphylococcus aureus is both a commensal of humans and a highly dangerous bacterial pathogen. EV production by S. aureus has been demonstrated and EVs shown to be cytolytic and play roles in immune activation. Studies of S. aureus EV composition have focused on their protein content and to date there are no reports describing the nucleic acid content of S. aureus EVs. S. aureus DNA and RNA are known to elicit an immune response however the mechanism(s) through which these nucleic acids gain access to the host cell is unknown. We hypothesize that EVs represent one mechanism through which S. aureus DNA/RNA is introduced into host cells. In this exploratory proposal we present preliminary data showing that RNA is located inside and associated with S. aureus EVs. We propose to identify the EV-associated RNA in EVs from five different S. aureus strains by RNA-sequencing, and to use that information to define the S. aureus EV-RNome – i.e. the RNA molecules found in EVs from all S. aureus strains. Defining the EV RNome will allow us to identify candidate RNA molecules for future analysis to determine their contribution to the immune stimulating properties of EVs.