Herpes simplex virus type 1 glycoprotein K is not essential for infectious virus production in actively replicating cells but is required for efficient envelopment and translocation of infectious virions from the cytoplasm to the extracellular space
Herpes simplex virus type 1 glycoprotein K is not essential for infectious virus production in actively replicating cells but is required for efficient envelopment and translocation of infectious virions from the cytoplasm to the extracellular space
复制标题
DOI：
10.1128/jvi.71.7.5012-5024.1997
复制
发表时间：
1997-07-01
影响因子：
5.4
通讯作者：
Kousoulas, KG
中科院分区：
文献类型：
--
作者：
Jayachandra, S;Baghian, A;Kousoulas, KG
我们对糖蛋白K（gK）缺失型单纯疱疹病毒1型[HSV-1]（科斯）Δ gK进行了表征，并将其与gK缺失型病毒HSV-1 F-gK β（L.哈钦森等人，J. Virol. 69：5401-5413，1995）。Δ gK和F-gK β突变病毒在感染后48 h在Vero细胞单层上产生小噬斑。F-gK β引起143 TK细胞的广泛融合，所述143 TK细胞对蜂毒肽敏感，蜂毒肽是gK诱导的细胞融合的特异性抑制剂，而Δ gK病毒不融合143 TK细胞。含有由F-gK β特异性的截短的gK基因的重组质粒未能拯救ICP 27-null病毒科斯（d27-1），而具有Δ gK缺失的质粒有效地拯救了d27-1病毒，Δ gK病毒产量在静止细胞中比在活跃复制的Vero细胞中低约100，000倍。Δ gK和F-gK β病毒原液在VK 302细胞上的噬斑效率相似，而F-gK β病毒原液在Vero细胞上的噬斑效率与Δ gK病毒相比降低近10，000倍。突变的Δ gK和F-gK β感染性病毒粒子在Vero和HEp-2细胞内积累，但不能移位到细胞外空间，Δ gK（衣壳在Vero细胞核中积累，但不在HEp-2细胞中积累。在Vero和HEp-2细胞的细胞质中可见包膜的Delta gK病毒体，并且在囊泡内的HEp-2细胞的细胞质中发现病毒衣壳。糖蛋白B、C、D和H在Delta gK感染的Vero细胞表面上以与KOS感染的Vero细胞相似的量表达。这些结果表明，gK参与核衣壳的形成，更重要的是参与感染性病毒体从细胞质到细胞外空间的移位。并且活跃复制的细胞可以部分地补偿DeltagK病毒的抑制，但不能补偿细胞排出缺陷。Delta gK和F-gICP病毒的比较表明，F-gK β病毒在Vero细胞中的低效病毒复制和空斑效率及其在143 TK（-）sells中的合胞体表型最有可能是由于截短的gK的表达。
We characterized the glycoprotein K (gK)-null herpes simplex virus type 1 [HSV-1] (KOS) Delta gK and compared it to the gK-null virus HSV-1 F-gK beta (L. Hutchinson et al., J. Virol. 69:5401-5413, 1995). Delta gK and F-gK beta mutant viruses produced small plaques on Vero cell monolayers at 48 h postinfertion. F-gK beta caused extensive fusion of 143TK cells that was sensitive to melittin, a specific inhibitor of gK-induced cell fusion, while Delta gK virus did not fuse 143TK cells, A recombinant plasmid containing the truncated gK gene specified by F-gK beta failed to rescue the ICP27-null virus KOS (d27-1), while a plasmid with the Delta gK deletion rescued the d27-1 virus efficiently, Delta gK virus yield was approximately 100,000-fold lower in stationary cells than in actively replicating Vero cells, The plaquing efficiencies of Delta gK and F-gK beta virus stocks on VK302 cells were similar, while the plaquing efficiency of F-gK beta virus stocks on Vero cells was reduced nearly 10,000-fold in comparison to that of Delta gK virus. Mutant Delta gK and F-gK beta infectious virions accumulated within Vero and HEp-2 cells but failed to translocate to extracellular spaces, Delta gK( capsids accumulated in the nuclei of Vero but not HEp-2 cells. Enveloped Delta gK virions were, visualized in the cytoplasms of both Vero and HEp-2 cells, and viral capsids were found in the cytoplasm of HEp-2, cells within vesicles. Glycoproteins B, C, D, and H were expressed on the surface of Delta gK-infected Vero cells in amounts similar Co those for KOS-infected Vero cells, These results indicate that gK is involved in nucleocapsid envelopment, and more importantly in the translocation of infectious virions from the cytoplasm to the extracellular spaces, and that actively replicating cells can partially compensate for the envelopment but not Tor the cellular egress deficiency; of the Delta gK virus. Comparison of Delta gK and F-gICP viruses suggests that the inefficient viral replication and plaquing efficiency of F-gK beta virus in Vero cells and its syncytial phenotype in 143TK(-) sells are most likely due to expression of a truncated gK.