Tran Robert K, Lieu Pauline T, Aguilar Santiago, Wagner Edward K, Bloom David C
Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Gainesville, FL 32610-0266, USA.
J Virol. 2002 Mar;76(5):2199-205. doi: 10.1128/jvi.76.5.2199-2205.2002.
While many herpes simplex virus (HSV) structural proteins are expressed with strict-late kinetics, the HSV virion protein 5 (VP5) is expressed as a "leaky-late" protein, such that appreciable amounts of VP5 are made prior to DNA replication. Our goal has been to determine if leaky-late expression of VP5 is a requirement for a normal HSV infection. It had been shown previously that recombinant viruses in which the VP5 promoter was replaced with promoters of other kinetic classes (including a strict late promoter) exhibited no alterations in replication kinetics or virus yields in vitro. In contrast, here we report that alterations in pathogenesis were observed when these recombinants were analyzed by experimental infection of mice. Following intracranial inoculation, a recombinant expressing VP5 from a strict-late promoter (U(L)38) exhibited an increased 50% lethal dose and a 10-fold decrease in virus yields in the central nervous system, while a recombinant expressing VP5 from an early (dUTPase) or another leaky-late (VP16) promoter exhibited wild-type neurovirulence. Moreover, following infection of the footpad, changing the expression kinetics of VP5 from leaky-late to strict-late resulted in 100-fold-less virus in the spinal ganglia during the acute infection than produced by either the parent virus or the rescued virus. These data indicate that the precise timing of appearance of the major capsid protein plays a role in the pathogenesis of HSV infections and that changing the expression kinetics has different effects in different cell types and tissues.
虽然许多单纯疱疹病毒(HSV)结构蛋白以严格晚期动力学表达,但HSV病毒体蛋白5(VP5)以“渗漏晚期”蛋白形式表达,以至于在DNA复制之前就产生了相当数量的VP5。我们的目标是确定VP5的渗漏晚期表达是否是HSV正常感染所必需的。先前已表明,将VP5启动子替换为其他动力学类别的启动子(包括严格晚期启动子)的重组病毒在体外复制动力学或病毒产量方面没有改变。相比之下,我们在此报告,当通过小鼠实验感染分析这些重组体时,观察到了发病机制的改变。颅内接种后,从严格晚期启动子(U(L)38)表达VP5的重组体在中枢神经系统中的50%致死剂量增加,病毒产量下降10倍,而从早期(dUTPase)或另一个渗漏晚期(VP16)启动子表达VP5的重组体表现出野生型神经毒力。此外,在足垫感染后,将VP5的表达动力学从渗漏晚期改变为严格晚期,导致急性感染期间脊髓神经节中的病毒比亲本病毒或拯救病毒产生的病毒少100倍。这些数据表明,主要衣壳蛋白出现的精确时间在HSV感染的发病机制中起作用,并且改变表达动力学在不同细胞类型和组织中有不同的影响。