Biological Functions and Cross-Kingdom Host Gene Regulation of Small RNAs in Lactobacillus plantarum-Derived Extracellular Vesicles.
Biological Functions and Cross-Kingdom Host Gene Regulation of Small RNAs in Lactobacillus plantarum-Derived Extracellular Vesicles.
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植物乳杆菌来源的细胞外囊泡中小RNA的生物学功能和跨界宿主基因调控
DOI：
10.3389/fmicb.2022.944361
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发表时间：
2022
影响因子：
5.2
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中科院分区：
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细胞外囊泡介导的microRNA转移是一种新的细胞间遗传传递模式。由微生物产生的胞外囊泡已被证明含有大量的生理活性分子，如蛋白质，脂质和RNA，它们可以被运输到宿主细胞并在界间信号传导和生理反应中起关键作用。在这项研究中，我们通过对植物乳杆菌来源的胞外囊泡（LDEVs）中的小RNA（sRNA）进行测序来鉴定sRNA，并使用定量逆转录-聚合酶链反应（RT-PCR）检测囊泡sRNA的表达水平，这证明了这些囊泡中存在microRNA大小的RNA（msRNA）。我们选择了一种高表达的msRNA sRNA 71进行进一步研究，预测了它在人类基因组中的潜在靶基因，并表明它可以易位到哺乳动物细胞中。随后，通过细胞蛋白质组学、蛋白质印迹和荧光素酶报告基因测定来探索该sRNA 71的生物学功能。结果表明，合成的sRNA 71模拟物转染HEK 293 T细胞后，可显著降低Tp 53的表达，并通过与Tp 53 mRNA的3′ UTR结合抑制其表达。总之，假设微生物来源的细胞外囊泡作为功能分子如sRNA的载体，其在调节微生物-宿主通讯中起重要作用。
Extracellular vesicle-mediated transfer of microRNAs is a novel mode of cell-to-cell genetic transmission. Extracellular vesicles produced by microbes have been shown to contain significant quantities of physiologically active molecules such as proteins, lipids, and RNA, which could be transported to host cells and play a key role in both inter-kingdom signaling and physiological responses. In this study, we identified sRNAs by sequencing small RNAs (sRNAs) from Lactobacillus plantarum-derived extracellular vesicles (LDEVs) and detected the expression levels of vesicular sRNAs using quantitative reverse transcription-polymerase chain reaction (RT-PCR), which demonstrated the presence of microRNA-sized RNAs (msRNAs) within these vesicles. We chose sRNA71, a highly expressed msRNA, for further investigation, predicted its potential target genes for the human genome, and indicated that it could be translocated into mammalian cells. The biological functions of this sRNA71 were subsequently explored through cellular proteomics, western blot, and luciferase reporter assay. According to the findings, transfection with synthetic sRNA71 mimics substantially reduced Tp53 expression in HEK293T cells and suppressed the gene expression through binding to the 3′ UTR of Tp53 mRNA. In conclusion, it is hypothesized that microbial-derived extracellular vesicles serve as carriers of functional molecules such as sRNAs, which play an essential role in regulating microbial-host communication.