Tal-Singer R, Podrzucki W, Lasner T M, Skokotas A, Leary J J, Fraser N W, Berger S L
The Wistar Institute, Philadelphia, Pennsylvania 19104-4268, USA.
J Virol. 1998 Feb;72(2):1252-61. doi: 10.1128/JVI.72.2.1252-1261.1998.
The detailed mechanism which governs the choice between herpes simplex virus (HSV) latency and reactivation remains to be elucidated. It is probable that altered expression of cellular factors in sensory neurons leads to induction of HSV gene expression resulting in reactivation. As an approach to identify novel cellular genes which are activated or repressed by stimuli that reactivate HSV from latency and hence may play a role in viral reactivation, RNA from explanted trigeminal ganglia (TG) was analyzed by differential display reverse transcription-PCR (DDRT-PCR). Nearly 50 cDNAs whose mRNA level was modified by the stress of explantation were isolated and sequenced. We present a listing of a spectrum of altered RNAs, including both known and unknown sequences. Five of those differentially displayed transcripts were identified as interferon-related murine TIS7 mRNA. These results were confirmed in both infected and uninfected ganglia by quantitative RNase protection assay and immunostaining. Alpha and beta interferons and interferon regulatory factor-1 (IRF-1) were also induced by explantation. In addition, we have identified sequences that correspond to IRF-1 consensus binding sites in both HSV type 1 origins of replication. Our findings suggest that physiological pathways that include these cellular factors may be involved in modulating HSV reactivation.
控制单纯疱疹病毒(HSV)潜伏与再激活之间选择的详细机制仍有待阐明。感觉神经元中细胞因子表达的改变可能导致HSV基因表达的诱导,从而引发再激活。作为一种识别由从潜伏状态重新激活HSV的刺激所激活或抑制的新型细胞基因的方法,通过差异显示逆转录PCR(DDRT-PCR)分析了从三叉神经节(TG)分离出的RNA。分离并测序了近50个其mRNA水平因外植应激而改变的cDNA。我们列出了一系列改变的RNA,包括已知和未知序列。其中五个差异显示的转录本被鉴定为与干扰素相关的小鼠TIS7 mRNA。通过定量核糖核酸酶保护测定和免疫染色在感染和未感染的神经节中证实了这些结果。外植也诱导了α和β干扰素以及干扰素调节因子-1(IRF-1)。此外,我们在1型HSV复制起点中都鉴定出了与IRF-1共有结合位点相对应的序列。我们的研究结果表明,包括这些细胞因子的生理途径可能参与调节HSV的再激活。