PDF(Pubmed)
Abstract：
Multiple functions are associated with HSV-1 latency associated transcript (LAT), including establishment of latency, virus reactivation, and antiapoptotic activity. LAT encodes two sncRNAs that are not miRNAs and previously it was shown that they have antiapoptotic activity in vitro. To determine if we can separate the antiapoptotic function of LAT from its latency-reactivation function, we deleted sncRNA1 and sncRNA2 sequences in HSV-1 strain McKrae, creating ΔsncRNA1&2 recombinant virus. Deletion of the sncRNA1&2 in ΔsncRNA1&2 virus was confirmed by complete sequencing of ΔsncRNA1&2 virus and its parental virus. Replication of ΔsncRNA1&2 virus in tissue culture or in the eyes of WT infected mice was similar to that of HSV-1 strain McKrae (LAT-plus) and dLAT2903 (LAT-minus) viruses. The levels of gB DNA in trigeminal ganglia (TG) of mice latently infected with ΔsncRNA1&2 virus was intermediate to that of dLAT2903 and McKrae infected mice, while levels of LAT in TG of latently infected ΔsncRNA1&2 mice was significantly higher than in McKrae infected mice. Similarly, the levels of LAT expression in Neuro-2A cells infected with ΔsncRNA1&2 virus was significantly higher than in McKrae infected cells. Reactivation in TG of ΔsncRNA1&2 infected mice was similar to that of McKrae and time of reactivation in both groups were significantly faster than dLAT2903 infected mice. However, levels of apoptosis in Neuro-2A cells infected with ΔsncRNA1&2 virus was similar to that of dLAT2903 and significantly higher than that of McKrae infected cells. Our results suggest that the antiapoptotic function of LAT resides within the two sncRNAs, which works independently of its latency-reactivation function and it has suppressive effect on LAT expression in vivo and in vitro.
摘要：
多种功能与HSV-1潜伏期相关转录物（LAT）相关，包括延迟的建立，病毒再激活,和抗凋亡活性。LAT编码两种不是miRNA的sncRNA,并且先前显示它们在体外具有抗凋亡活性。为了确定我们是否可以将LAT的抗凋亡功能与其潜伏期再激活功能分开，我们在HSV-1株McKrae中删除了sncRNA1和sncRNA2序列，产生ΔsncRNA1&2重组病毒。通过对ΔsncRNA1&2病毒及其亲本病毒进行完全测序,确认ΔsncRNA1&2病毒中sncRNA1&2的缺失。ΔsncRNA1＆2病毒在受WT感染的小鼠的组织培养或眼中的复制与HSV-1株McKrae（LAT-plus）和dLAT2903（LAT-负）病毒相似。潜伏感染ΔsncRNA1和2病毒的小鼠的三叉神经节（TG）中gBDNA的水平介于dLAT2903和McKrae感染的小鼠的水平，而潜伏感染的ΔsncRNA1和2小鼠的TG中LAT的水平明显高于McKrae感染的小鼠。同样，用ΔsncRNA1和2病毒感染的Neuro-2A细胞中的LAT表达水平明显高于McKrae感染的细胞。ΔsncRNA1＆2感染的小鼠的TG再激活与McKrae相似，两组的再激活时间均明显快于dLAT2903感染的小鼠。然而,用ΔsncRNA1和2病毒感染的Neuro-2A细胞的凋亡水平与dLAT2903相似，并显着高于McKrae感染的细胞。我们的结果表明，LAT的抗凋亡功能存在于两个sncRNA中，其工作独立于其潜伏期再激活功能，并且在体内和体外对LAT表达具有抑制作用。
