Inman M, Perng G C, Henderson G, Ghiasi H, Nesburn A B, Wechsler S L, Jones C
Department of Veterinary and Biomedical Sciences, University of Nebraska, Lincoln, Nebraska 68583-0905, USA.
J Virol. 2001 Apr;75(8):3636-46. doi: 10.1128/JVI.75.8.3636-3646.2001.
The latency-associated transcript (LAT) is the only abundant herpes simplex virus type 1 (HSV-1) transcript expressed during latency. In the rabbit eye model, LAT null mutants do not reactivate efficiently from latency. We recently demonstrated that the LAT null mutant dLAT2903 induces increased levels of apoptosis in trigeminal ganglia of infected rabbits compared to LAT+ strains (G.-C. Perng, C. Jones, J. Ciacci-Zarella, M. Stone, G. Henderson, A. Yokht, S. M. Slanina, F. M. Hoffman, H. Ghiasi, A. B. Nesburn, and C. S. Wechsler, Science 287:1500-1503, 2000). The same study also demonstrated that a plasmid expressing LAT nucleotides 301 to 2659 enhanced cell survival of transfected cells after induction of apoptosis. Consequently, we hypothesized that LAT enhances spontaneous reactivation in part, because it promotes survival of infected neurons. Here we report on the ability of plasmids expressing different portions of the 5' end of LAT to promote cell survival after induction of apoptosis. A plasmid expressing the first 1.5 kb of LAT (LAT nucleotides 1 to 1499) promoted cell survival in neuro-2A (mouse neuronal) and CV-1 (monkey fibroblast) cells. A plasmid expressing just the first 811 nucleotides of LAT promoted cell survival less efficiently. Plasmids expressing the first 661 nucleotides or less of LAT did not promote cell survival. We previously showed that a mutant expressing just the first 1.5 kb of LAT has wild-type spontaneous reactivation in rabbits, and a mutant expressing just the first 811 nucleotides of LAT has a reactivation frequency higher than that of dLAT2903 but lower than that of wild-type virus. In addition, mutants reported here for the first time, expressing just the first 661 or 76 nucleotides of LAT, had spontaneous reactivation indistinguishable from that of the LAT null mutant dLAT2903. In summary, these studies provide evidence that there is a functional relationship between the ability of LAT to promote cell survival and its ability to enhance spontaneous reactivation.
潜伏期相关转录本(LAT)是单纯疱疹病毒1型(HSV-1)在潜伏期间唯一大量表达的转录本。在兔眼模型中,LAT基因缺失突变体不能有效地从潜伏状态重新激活。我们最近证明,与LAT+毒株相比,LAT基因缺失突变体dLAT2903在受感染兔子的三叉神经节中诱导的细胞凋亡水平增加(G.-C. 彭、C. 琼斯、J. 恰奇-扎雷拉、M. 斯通、G. 亨德森、A. 约克特、S. M. 斯拉尼纳、F. M. 霍夫曼、H. 贾西亚、A. B. 内斯本和C. S. 韦克斯勒,《科学》287:1500 - 1503,2000)。同一研究还表明,表达LAT核苷酸301至2659的质粒在诱导细胞凋亡后可提高转染细胞的存活率。因此,我们推测LAT部分地增强了自发重新激活,因为它促进了受感染神经元的存活。在此,我们报告了表达LAT 5'端不同部分的质粒在诱导细胞凋亡后促进细胞存活的能力。一个表达LAT前1.5 kb(LAT核苷酸1至1499)的质粒在Neuro-2A(小鼠神经元)和CV-1(猴成纤维细胞)细胞中促进了细胞存活。一个仅表达LAT前811个核苷酸的质粒促进细胞存活的效率较低。表达LAT前661个核苷酸或更少的质粒不能促进细胞存活。我们之前表明,一个仅表达LAT前1.5 kb的突变体在兔子中具有野生型自发重新激活能力,一个仅表达LAT前811个核苷酸的突变体的重新激活频率高于dLAT2903,但低于野生型病毒。此外,这里首次报道的仅表达LAT前661或76个核苷酸的突变体,其自发重新激活与LAT基因缺失突变体dLAT2903没有区别。总之,这些研究提供了证据,表明LAT促进细胞存活的能力与其增强自发重新激活的能力之间存在功能关系。