STRUCTURE OF THE HERPES-SIMPLEX VIRUS CAPSID - EFFECTS OF EXTRACTION WITH GUANIDINE-HYDROCHLORIDE AND PARTIAL RECONSTITUTION OF EXTRACTED CAPSIDS
STRUCTURE OF THE HERPES-SIMPLEX VIRUS CAPSID - EFFECTS OF EXTRACTION WITH GUANIDINE-HYDROCHLORIDE AND PARTIAL RECONSTITUTION OF EXTRACTED CAPSIDS
复制标题
DOI：
10.1128/jvi.65.2.613-620.1991
复制
发表时间：
1991-02-01
影响因子：
5.4
通讯作者：
BROWN, JC
中科院分区：
文献类型：
--
作者：
NEWCOMB, WW;BROWN, JC
从单纯疱疹病毒1型感染的细胞中纯化病毒B衣壳，并用2.0 M盐酸胍（GuHCl）体外提取。 十二烷基硫酸钠-聚丙烯酰胺凝胶分析表明，提取导致超过95%的衣壳蛋白VP 22 a和VP 26的去除，而VP 5（主要衣壳蛋白）、VP 19和VP 23的损失很小（小于10%）。 提取的衣壳的电子显微镜分析显示，五邻体和在B衣壳的空腔内发现的物质（主要是VP 22 a）几乎定量地被去除，但提取的衣壳在其他方面保持结构完整。 几乎没有六邻体丢失;衣壳直径没有受到很大影响;其二十面体对称性仍然很明显。 结果表明，VP 19和VP 23都不能构成衣壳五邻体。 与六邻体一样，五邻体最有可能由VP 5组成。 当B衣壳用2.0 M GuHCl处理，然后透析除去GuHCl时，通过蔗糖密度梯度超离心分离两条带的病毒物质。 两者中迁移更快的衣壳缺乏五邻体和VP 22 a，但具有完整的VP 26。 因此，VP 26必须在透析期间与提取的衣壳重新结合。 更缓慢迁移的条带由直径约60 nm的环形结构组成，其完全由VP 22 a组成。 后者的结构非常类似于天然B衣壳腔中常见的环面状浓缩物。 结果表明单纯疱疹病毒1型B衣壳和沙门氏菌噬菌体P22的前衣壳之间的相似性。 两者都含有内部蛋白（HSV-1 B衣壳中的VP 22 a和噬菌体P22中的gp 8或“支架”蛋白），其可以用GuHCl在体外提取并且不存在于成熟病毒体中。
Viral B capsids were purified from cells infected with herpes simplex virus type 1 and extracted in vitro with 2.0 M guanidine hydrochloride (GuHC1). Sodium dodecyl sulfate-polyacrylamide gel analyses demonstrated that extraction resulted in the removal of greater than 95% of capsid proteins VP22a and VP26 while there was only minimal (less than 10%) loss of VP5 (the major capsid protein), VP19, and VP23. Electron microscopic analysis of extracted capsids revealed that the pentons and the material found inside the cavity of B capsids (primarily VP22a) were removed nearly quantitatively, but extracted capsids remained otherwise structurally intact. Few, if any, hexons were lost; the capsid diameter was not greatly affected; and its icosahedral symmetry was still clearly evident. The results demonstrate that neither VP19 nor VP23 could constitute the capsid pentons. Like the hexons, the pentons are most likely composed of VP5. When B capsids were treated with 2.0 M GuHCl and then dialyzed to remove GuHCl, two bands of viral material were separated by sucrose density gradient ultracentrifugation. The more rapidly migrating of the two consisted of capsids which lacked pentons and VP22a but had a full complement of VP26. Thus, VP26 must have reassociated with extracted capsids during dialysis. The more slowly migrating band consisted of torus-shaped structures approximately 60 nm in diameter which were composed entirely of VP22a. These latter structures closely resembled torus-shaped condensates often seen in the cavity of native B capsids. The results suggest a similarity between herpes simplex virus type 1 B capsids and procapsids of Salmonella bacteriophage P22. Both contain an internal protein (VP22a in the case of HSV-1 B capsids and gp8 or "scaffolding" protein in phage P22) that can be extracted in vitro with GuHCl and that is absent from mature virions.