Winterling Carla, Koch Manuel, Koeppel Max, Garcia-Alcalde Fernando, Karlas Alexander, Meyer Thomas F
Department of Molecular Biology; Max Planck Institute for Infection Biology; Berlin, Germany.
RNA Biol. 2014;11(1):66-75. doi: 10.4161/rna.27504. Epub 2013 Dec 20.
A growing body of evidence suggests the non-protein coding human genome is of vital importance for human cell function. Besides small RNAs, the diverse class of long non-coding RNAs (lncRNAs) recently came into focus. However, their relevance for infection, a major evolutionary driving force, remains elusive. Using two commercially available microarray systems, namely NCode™ and Sureprint™ G3, we identified differential expression of 42 ncRNAs during influenza A virus (IAV) infection in human lung epithelial cells. This included several classes of lncRNAs, including large intergenic ncRNAs (lincRNAs). As analyzed by qRT-PCR, expression of one lincRNA, which we termed virus inducible lincRNA (VIN), is induced by several IAV strains (H1N1, H3N2, H7N7) as well as vesicular stomatitis virus. However, we did not observe an induction of VIN by influenza B virus, treatment with RNA mimics, or IFNβ. Thus, VIN expression seems to be a specific response to certain viral infections. RNA fractionation and RNA-FISH experiments revealed that VIN is localized to the host cell nucleus. Most importantly, we show that abolition of VIN by RNA interference restricts IAV replication and viral protein synthesis, highlighting the relevance of this lincRNA for productive IAV infection. Our observations suggest that viral pathogens interfere with the non-coding portion of the human genome, thereby guaranteeing their successful propagation, and that the expression of VIN correlates with their virulence. Consequently, our study provides a novel approach for understanding virus pathogenesis in greater detail, which will enable future design of new antiviral strategies targeting the host's non-protein coding genome.
越来越多的证据表明，非蛋白质编码人类基因组对人类细胞功能至关重要。除了小RNA外，最近，种类多样的长链非编码RNA（lncRNA）也受到了关注。然而，它们与感染（一种主要的进化驱动力）之间的相关性仍不清楚。我们使用两种市售微阵列系统，即NCode™和Sureprint™ G3，鉴定了人类肺上皮细胞在甲型流感病毒（IAV）感染期间42种非编码RNA的差异表达。这包括几类lncRNA，包括大基因间非编码RNA（lincRNA）。通过qRT-PCR分析，我们将一种lincRNA命名为病毒诱导性lincRNA（VIN），它可被几种IAV毒株（H1N1、H3N2、H7N7）以及水疱性口炎病毒诱导表达。然而，我们未观察到乙型流感病毒、RNA模拟物处理或IFNβ诱导VIN表达。因此，VIN表达似乎是对某些病毒感染的特异性反应。RNA分级分离和RNA-FISH实验表明，VIN定位于宿主细胞核。最重要的是，我们发现通过RNA干扰消除VIN可限制IAV复制和病毒蛋白合成，突出了这种lincRNA对IAV有效感染的相关性。我们的观察结果表明，病毒病原体干扰人类基因组的非编码部分，从而确保其成功传播，并且VIN的表达与其毒力相关。因此，我们的研究为更详细地理解病毒发病机制提供了一种新方法，这将有助于未来设计针对宿主非蛋白质编码基因组的新型抗病毒策略。
