Evaluation of the primary effect of brefeldin A treatment upon herpes simplex virus assembly
Evaluation of the primary effect of brefeldin A treatment upon herpes simplex virus assembly
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DOI：
10.1099/0022-1317-82-7-1561
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发表时间：
2001-07-01
影响因子：
3.8
通讯作者：
Wilson, DW
中科院分区：
文献类型：
--
作者：
Dasgupta, A;Wilson, DW
将药物brefeldin A （BFA）添加到被1型单纯疱疹病毒（HSV）感染的细胞中，已知会导致颗粒组装中出现复杂的缺陷模式。经BFA处理的感染细胞在核周包膜病毒粒子和非包膜（“裸”）细胞质衣壳上积累，很难根据HSV的组装途径和BFA对分泌器官的已知影响来解释这些数据。由于BFA是一种细胞毒性药物，并且早期的研究通常检查BFA长期孵育对感染细胞的影响，因此假设该药物可能对HSV组装具有多效性和间接作用。为了验证这一点，使用了HSV同步组装试验，其中细胞被病毒突变体tsProt，A感染并保持在39°c以诱导原囊体群体的可逆积累。首先加入BFA，然后将这些细胞转移到31度，使积累的原壳成熟3小时，可以测试短期BFA处理仅对原壳成熟下游的HSV组装事件的影响。在这些条件下，原囊体成熟并正常包装病毒基因组，但仍未被包膜，不能离开细胞核。由此可见，BFA对HSV复制的主要作用是抑制核膜上的出芽。
Addition of the drug brefeldin A (BFA) to cells infected by herpes simplex virus (HSV) type 1 is known to result in a complex pattern of defects in particle assembly. BFA-treated, infected cells accumulate perinuclear enveloped virions and non-enveloped ('naked') cytoplasmic capsids, and it has been difficult to interpret these data in terms of the assembly pathway of HSV and the known effects of BFA on the secretory apparatus. Since BFA is a cytotoxic drug, and earlier studies commonly examined the effects of long-term BFA incubations on infected cells, it was hypothesized that the drug could have pleiotropic and indirect effects on HSV assembly. To test this, use was made of an HSV synchronized assembly assay, in which cells are infected with the virus mutant tsProt,A and maintained at 39 degreesC to induce reversible accumulation of a population of procapsids. By first adding BFA and then shifting these cells to 31 degreesC for 3 h to allow the accumulated procapsids to mature, it was possible to test the effect of short-term BFA treatment on only those HSV assembly events that are downstream of procapsid maturation. Under these conditions, it was found that procapsids matured and packaged the viral genome normally, but remained non-enveloped and failed to exit the nucleus. It is concluded that the primary effect of BFA on HSV replication is to inhibit budding at the inner nuclear membrane.