Wu Zetang, Zhu Yali, Bisaro David M, Parris Deborah S
Graduate Program in Molecular, Cellular, and Developmental Biology, Ohio State University, Columbus, Ohio 43210, USA.
J Virol. 2009 Jul;83(13):6652-63. doi: 10.1128/JVI.00260-09. Epub 2009 Apr 15.
RNA-induced silencing is a potent innate antiviral defense strategy in plants, and suppression of silencing is a hallmark of pathogenic plant viruses. However, the impact of silencing as a mammalian antiviral defense mechanism and the ability of mammalian viruses to suppress silencing in natural host cells have remained controversial. The ability of herpes simplex virus type 1 (HSV-1) to suppress silencing was examined in a transient expression system that employed an imperfect hairpin to target degradation of transcripts encoding enhanced green fluorescent protein (EGFP). HSV-1 infection suppressed EGFP-specific silencing as demonstrated by increased EGFP mRNA levels and an increase in the EGFP mRNA half-life. The increase in EGFP mRNA stability occurred despite the well-characterized host macromolecular shutoff functions of HSV-1 that globally destabilize mRNAs. Moreover, mutant viruses defective in these functions increased the stability of EGFP mRNA even more than did the wild-type virus in silenced cells compared to results in control cells. The importance of RNA silencing to HSV-1 replication was confirmed by a significantly enhanced virus burst size in cells in which silencing was knocked down with small inhibitory RNAs directed to Argonaute 2, an integral component of the silencing complex. Given that HSV-1 encodes several microRNAs, it is possible that a dynamic equilibrium exists between silencing and silencing suppression that is capable of modulating viral gene expression to promote replication, to evade host defenses, and/or to promote latency.
RNA诱导的沉默是植物中一种强大的先天性抗病毒防御策略,而沉默的抑制是致病植物病毒的一个标志。然而,沉默作为哺乳动物抗病毒防御机制的影响以及哺乳动物病毒在天然宿主细胞中抑制沉默的能力一直存在争议。在一个瞬时表达系统中检测了1型单纯疱疹病毒(HSV-1)抑制沉默的能力,该系统使用一个不完美的发夹来靶向降解编码增强型绿色荧光蛋白(EGFP)的转录本。HSV-1感染抑制了EGFP特异性沉默,这表现为EGFP mRNA水平的增加以及EGFP mRNA半衰期的延长。尽管HSV-1具有已被充分表征的宿主大分子关闭功能,可使mRNA整体不稳定,但EGFP mRNA稳定性仍有所增加。此外,与对照细胞相比,在沉默细胞中,这些功能有缺陷的突变病毒比野生型病毒更能增加EGFP mRNA的稳定性。通过用针对沉默复合物的一个组成部分AGO2的小干扰RNA敲低沉默,显著增强了病毒爆发大小,从而证实了RNA沉默对HSV-1复制的重要性。鉴于HSV-1编码几种微小RNA,沉默与沉默抑制之间可能存在一种动态平衡,这种平衡能够调节病毒基因表达以促进复制、逃避宿主防御和/或促进潜伏。