Characterization of Envelopment Process of viral Nucleocapsids in Herpes Simplex Virus-Infected Cells

我们定量研究了单纯疱疹病毒1型（HSV-1）在Vero细胞内的增殖。氯化铵对HSV-1增殖抑制作用的表征揭示了病毒颗粒的形成（即，病毒核衣壳的分解）发生在细胞内酸性区室中，并且区室中的酸性对于分解是必需的。为了检测该试剂对病毒刺突蛋白的合成和转运的影响，表征了在HSV-1感染的细胞表面上吸附致敏红细胞的能力的外观。动力学的外观表明，刺突蛋白的合成和运输不受agent.Since最酸性和最剂敏感的隔间细胞内的细胞器之间是已知的高尔基复合体，我们研究了高尔基复合体的作用，以及在HSV-1感染的细胞中的感染性子代病毒的形成。由于Orefeldin A（BFA）最近被报道引起高尔基复合体的解体，我们表征了BFA对HSV-1增殖的影响，表明（1）存在两种不同的（i.例如，（2）感染性子代病毒完全由药物敏感途径产生，这些结果表明高尔基复合体在HSV-1感染细胞中的增殖过程中起着重要作用，导致感染性子代病毒的形成。

We have quantitatively studied an intracellular multiplication of herpes simplex virus type 1 (HSV-1) in Vero cells. Characterization of the inhibition of the multiplication of HSV-1 by ammonium chloride revealed that the formation of the virus particles (i, e., the envelopment of viral nucleocapsids) takes place in the intracellular acidic compartments and that the acidity in the compartments is essential for the envelopment. To examine the effect of the agent on the synthesis and transport of spike proteins of virus, the appearance of ability to adsorb the sensitized erythrocytes on the HSV-1-infected cell surface was characterized. The kinetics of appearance indicated that both synthesis and transport of spike proteins are not affected by the agent.Since the most acidic and the most agent-sensitive compartment among intracellular organelles is known to be the Golgi complex, we examined the role of the Golgi complex in the envelopment as well as in the formation of infectious progeny virus in the HSV-1-infected cells. Since Orefeldin A (BFA) is recently reported to cause disassembly of the Golgi complex, we characterized the effect of BFA on the HSV-1 multiplication, showing that (1) there are two different (i. e., agent-sensitive and -insensitive) pathways for the formation of virus particles in the HSV-1-infected cells and that (2) the infectious progeny virus arises exclusively from the agent-sensitive pathway.These results indicated the crucial role of the Golgi complex in the envelopment of HSV-1 that lead to the formation of the infectious progeny virus.