Abstract:
Exosomes may function as multifactorial mediators of cell-to-cell communication, playing crucial roles in both physiological and pathological processes. Exosomes released from virus-infected cells may contain RNA and proteins facilitating infection spread. The purpose of our study was to analyze how the small RNA content of exosomes is affected by infection with the influenza A virus (IAV). Exosomes were isolated by ultracentrifugation after hemadsorption of virions and their small RNA content was identified using high-throughput sequencing. As compared to mock-infected controls, 856 RNA transcripts were significantly differentially expressed in exosomes from IAV-infected cells, including fragments of 458 protein-coding (pcRNA), 336 small, 28 long intergenic non-coding RNA transcripts, and 33 pseudogene transcripts. Upregulated pcRNA species corresponded mainly to proteins associated with translation and antiviral response, and the most upregulated among them were RSAD2, CCDC141 and IFIT2. Downregulated pcRNA species corresponded to proteins associated with the cell cycle and DNA packaging. Analysis of differentially expressed pseudogenes showed that in most cases, an increase in the transcription level of pseudogenes was correlated with an increase in their parental genes. Although the role of exosome RNA in IAV infection remains undefined, the biological processes identified based on the corresponding proteins may indicate the roles of some of its parts in IAV replication.
摘要:
外泌体可以作为细胞间通讯的多因素介质,在生理和病理过程中起着至关重要的作用。从病毒感染的细胞释放的外来体可能含有促进感染传播的RNA和蛋白质。我们研究的目的是分析外泌体的小RNA含量如何受到甲型流感病毒(IAV)感染的影响。病毒粒子吸附后,通过超速离心分离外泌体,并使用高通量测序鉴定其小RNA含量。与模拟感染的对照相比,856个RNA转录本在来自IAV感染细胞的外泌体中显著差异表达,包括458蛋白质编码(pcRNA)的片段,336小,28个长基因间非编码RNA转录本,和33个假基因转录本.上调的pcRNA种类主要对应于与翻译和抗病毒反应相关的蛋白质,其中上调幅度最大的是RSAD2、CCDC141和IFIT2。下调的pcRNA种类对应于与细胞周期和DNA包装相关的蛋白质。差异表达假基因的分析表明,在大多数情况下,假基因转录水平的增加与其亲本基因的增加有关。尽管外泌体RNA在IAV感染中的作用尚不明确,基于相应蛋白质鉴定的生物过程可能表明其某些部分在IAV复制中的作用。