Cavalcoli J D, Baghian A, Homa F L, Kousoulas K G
Department of Veterinary Microbiology and Parasitology, School of Veterinary Medicine, Louisiana State University, Baton Rouge 70803.
Virology. 1993 Nov;197(1):23-34. doi: 10.1006/viro.1993.1563.
Herpes simplex virus type 1 (HSV-1) mutant tsZ47 was reported to be temperature sensitive for virus growth and transport of viral glycoproteins to the cell surface and to contain two different mutations (B. A. Pancake, D. P. Aschman, and P. A. Schaffer, (1983) J. Virol. 47,568-585). However, we found that similar amounts of glycoproteins B, C and H were expressed at the cell surface at the permissive and non-permissive temperatures and in addition, tsZ47 virus contained only a single mutation. UL28-null virus, gC delta 7B, failed to complement tsZ47 in mixed infections and tsZ47 replicated in UL28 but not gB transformed cell lines. A ts lesion of tsZ47 was mapped within a 1333 bp region of the UL28 gene by marker-rescue using overlapping DNA fragments. DNA sequencing identified a C to T transversion resulting in an R to W amino acid change at UL28 amino acid position 531. Southern Blot analysis and transmission electron microscopy demonstrated that tsZ47, is defective in cleavage and encapsidation of viral DNA. Mutant virus ts1203 (C. Addison, F.J. Rixon, and V.G. Preston, (1990) J. Gen. Virol. 71, 2377-2384) that contains a mutation in the 5' end of UL28, complemented tsZ47 in mixed infections. This suggests that allelic complementation may be occurring and UL28 may encode a protein with independently functioning domains, or that it participates in a multimer.
据报道，单纯疱疹病毒1型（HSV-1）突变体tsZ47对病毒生长以及病毒糖蛋白向细胞表面的转运具有温度敏感性，并且含有两种不同的突变（B. A. Pancake、D. P. Aschman和P. A. Schaffer，（1983年）《病毒学杂志》47卷，568 - 585页）。然而，我们发现，在允许温度和非允许温度下，细胞表面表达的糖蛋白B、C和H的量相似，此外，tsZ47病毒仅含有一个单一突变。UL28缺失病毒gC delta 7B在混合感染中无法互补tsZ47，并且tsZ47在UL28但不在gB转化的细胞系中复制。通过使用重叠DNA片段的标记拯救，将tsZ47的一个温度敏感损伤定位在UL28基因的一个1333 bp区域内。DNA测序确定了一个C到T的颠换，导致UL28氨基酸位置531处的R到W氨基酸变化。Southern印迹分析和透射电子显微镜表明，tsZ47在病毒DNA的切割和包装方面存在缺陷。在UL28 5'端含有一个突变的突变病毒ts1203（C. Addison、F.J. Rixon和V.G. Preston，（1990年）《普通病毒学杂志》71卷，2377 - 2384页）在混合感染中互补了tsZ47。这表明可能发生了等位基因互补，并且UL28可能编码一种具有独立功能结构域的蛋白质，或者它参与了一个多聚体。
