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首页> 外文期刊>Journal of Extracellular Vesicles >The HIV Nef protein modulates cellular and exosomal miRNA profiles in human monocytic cells
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The HIV Nef protein modulates cellular and exosomal miRNA profiles in human monocytic cells

机译:HIV Nef蛋白调节人单核细胞中的细胞和外泌体miRNA谱

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IntroductionThe HIV Nef protein is a multifunctional virulence factor that perturbs intracellular membranes and signalling and is secreted into exosomes. While Nef-containing exosomes have a distinct proteomic profile, no comprehensive analysis of their miRNA cargo has been carried out. Since Nef functions as a viral suppressor of RNA interference and disturbs the distribution of RNA-induced silencing complex proteins between cells and exosomes, we hypothesized that it might also affect the export of miRNAs into exosomes.MethodExosomes were purified from human monocytic U937 cells that stably expressed HIV-1 Nef. The RNA from cells and exosomes was profiled for 667 miRNAs using a Taqman Low Density Array. Selected miRNAs and their mRNA targets were validated by quantitative RT-PCR. Bioinformatics analyses were used to identify targets and predict pathways.ResultsNef expression affected a significant fraction of miRNAs in U937 cells. Our analysis showed 47 miRNAs to be selectively secreted into Nef exosome...
机译:简介HIV Nef蛋白是一种多功能毒力因子,会干扰细胞内膜和信号传导并分泌到外泌体中。虽然含Nef的外泌体具有明显的蛋白质组学特征,但尚未对其miRNA货物进行全面分析。由于Nef起到RNA干扰的病毒抑制作用并干扰RNA诱导的沉默复合蛋白在细胞和外泌体之间的分布,因此我们推测它可能也会影响miRNA向外泌体的输出。方法外泌体是从人单核U937细胞中稳定纯化的表达的HIV-1 Nef。使用Taqman低密度阵列分析了来自细胞和外泌体的RNA,分析了667个miRNA。所选的miRNA及其mRNA靶标通过定量RT-PCR验证。通过生物信息学分析来鉴定靶标并预测途径。结果Nef表达影响了U937细胞中大部分miRNA。我们的分析表明,有47种miRNA被选择性地分泌到Nef外泌体中。

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