Priola S A, Gustafson D P, Wagner E K, Stevens J G
Department of Microbiology and Immunology, University of California, Los Angeles School of Medicine 90024.
J Virol. 1990 Oct;64(10):4755-60. doi: 10.1128/JVI.64.10.4755-4760.1990.
Pseudorabies virus (PRV) is a porcine herpesvirus that establishes latent infections in trigeminal ganglia. To determine whether PRV expresses any transcripts that could play a role in latency, the trigeminal ganglia of 14 pigs previously inoculated through the nose and latently infected with PRV(Ka) were assayed by in situ nucleic acid hybridization for the presence of PRV-specific RNA. Hybridizations employing probes encompassing the entire viral genome revealed that an area extending from 0.64 to 0.82 map units was transcriptionally active. The DNA probe that most consistently detected transcripts was BamHI-8, a fragment which contains the gene for the immediate-early protein. With this probe, ganglia from 10 (71%) of 14 pigs scored positive for PRV RNA, although only 1 (8%) of 12 of the ganglia from the opposite side reactivated virus after explanation and culture of latently infected trigeminal ganglia. The RNA was transcribed from the strand opposite to that coding for the immediate-early protein; the signal was neuronally localized, with dense nuclear accumulation accompanied by variable numbers of grains over the cytoplasm. Northern RNA blot analysis showed that a discrete set of poly(A)- PRV transcripts were present in latently infected trigeminal ganglia. Additional in situ nucleic acid hybridization analysis revealed that the 3' limit of the transcriptionally active area was located in a 1.2-kilobase fragment upstream and adjacent to the 5' end of the immediate-early protein RNA, whereas the 5' limit was as much as 4.9 kilobases downstream from the 3' end of this RNA. PRV therefore expresses latent-phase transcripts that, although similar in many respects to latent-phase transcripts reported for other herpesviruses, have some unique properties.
伪狂犬病病毒(PRV)是一种猪疱疹病毒,可在三叉神经节中建立潜伏感染。为了确定PRV是否表达任何可能在潜伏中起作用的转录本,对14头先前经鼻接种并潜伏感染PRV(Ka)的猪的三叉神经节进行了原位核酸杂交检测,以检测PRV特异性RNA的存在。使用涵盖整个病毒基因组的探针进行杂交显示,从0.64到0.82图谱单位的区域具有转录活性。最能持续检测到转录本的DNA探针是BamHI-8,该片段包含立即早期蛋白的基因。用该探针检测,14头猪中有10头(71%)的神经节PRV RNA检测呈阳性,尽管在对潜伏感染的三叉神经节进行解剖和培养后,12对神经节中只有1对(8%)重新激活了病毒。RNA是从与编码立即早期蛋白的链相反的链转录而来的;信号定位于神经元,细胞核密集积聚,细胞质上有数量不等的颗粒。Northern RNA印迹分析表明,潜伏感染的三叉神经节中存在一组离散的多聚腺苷酸(poly(A))-PRV转录本。进一步的原位核酸杂交分析表明,转录活性区域的3'端位于立即早期蛋白RNA 5'端上游和相邻的一个1.2千碱基片段中,而5'端则在该RNA 3'端下游多达4.9千碱基处。因此,PRV表达潜伏阶段转录本,尽管在许多方面与其他疱疹病毒报道的潜伏阶段转录本相似,但具有一些独特的特性。