Rock D L, Nesburn A B, Ghiasi H, Ong J, Lewis T L, Lokensgard J R, Wechsler S L
Department of Veterinary Science, North Dakota State University, Fargo 50105.
J Virol. 1987 Dec;61(12):3820-6. doi: 10.1128/JVI.61.12.3820-3826.1987.
Using a combination of in situ hybridization and Northern (RNA) blot analysis, we investigated herpes simplex virus type 1 (HSV-1) transcriptional activity in an ocular rabbit model of HSV-1 latency. Radioactively labeled cloned fragments, representing virtually the entire HSV-1 genome, were individually hybridized to RNA in sections of trigeminal ganglia taken from rabbits during the latent phase of infection with HSV-1 (McKrae). Our results suggest that two discrete latency-related RNAs (LR-RNAs) may be present. The LR-RNAs were localized mainly in the nuclei of neurons. The more abundant LR-RNA was detected in approximately 3% of all neurons examined and was designated major LR-RNA. The other LR-RNA, designated minor LR-RNA, was detected in approximately 0.3% of neurons from latently infected rabbits. The genes for the LR-RNAs mapped in the vicinity of the immediate-early gene ICP0 (also designated IE110). The gene for the major LR-RNA partially overlapped the left (3') end of the ICP0 gene. In situ hybridization with single-stranded RNA probes showed that this LR-RNA was of complementary sense to that of ICP0 mRNA. Northern blot analysis gave an approximate size for this LR-RNA of 1.8 to 2.2 kilobases. The minor LR-RNA mapped to or near the right (5') end of the ICP0 gene. The detection of LR-RNAs suggests the possibility that these RNAs or their products may play significant roles in the initiation and/or maintenance of HSV-1 latency.
我们运用原位杂交和Northern(RNA)印迹分析相结合的方法,研究了单纯疱疹病毒1型(HSV-1)在兔眼部潜伏模型中的转录活性。用放射性标记的克隆片段(几乎代表了整个HSV-1基因组),分别与处于HSV-1潜伏感染期(McKrae)的兔三叉神经节切片中的RNA进行杂交。我们的结果表明,可能存在两种不同的潜伏相关RNA(LR-RNA)。LR-RNA主要定位于神经元细胞核中。在所检测的所有神经元中,约3%的神经元检测到丰度较高的LR-RNA,将其命名为主要LR-RNA。另一种LR-RNA,命名为次要LR-RNA,在潜伏感染兔的约0.3%的神经元中检测到。LR-RNA的基因定位于立即早期基因ICP0(也称为IE110)附近。主要LR-RNA的基因部分重叠于ICP0基因的左侧(3')末端。用单链RNA探针进行原位杂交表明,这种LR-RNA与ICP0 mRNA的意义互补。Northern印迹分析得出这种LR-RNA的大小约为1.8至2.2千碱基。次要LR-RNA定位于ICP0基因的右侧(5')末端或其附近。LR-RNA的检测提示了这些RNA或其产物可能在HSV-1潜伏的起始和/或维持中发挥重要作用的可能性。