MUTATIONS IN HERPES-SIMPLEX VIRUS TYPE-1 GENES ENCODING VP5 AND VP23 ABROGATE CAPSID FORMATION AND CLEAVAGE OF REPLICATED DNA
MUTATIONS IN HERPES-SIMPLEX VIRUS TYPE-1 GENES ENCODING VP5 AND VP23 ABROGATE CAPSID FORMATION AND CLEAVAGE OF REPLICATED DNA
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DOI：
10.1128/jvi.67.3.1357-1364.1993
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发表时间：
1993-03-01
影响因子：
5.4
通讯作者：
PERSON, S
中科院分区：
文献类型：
--
作者：
DESAI, P;DELUCA, NA;PERSON, S
单纯疱疹病毒1型衣壳由七种衣壳蛋白组成，称为VP 5、VP 19 c、VP 21、VP 22 a、VP 23、VP 24和VP 26。主要衣壳蛋白VP 5由基因UL 19编码。UL 18，其转录物与VP 5的转录物3'共末端，特异性地表达衣壳蛋白VP 23。已经分离出用科斯的BglII N（UL 19）或EcoRI G（UL 16至UL 21）片段转化的Vero细胞系。这些细胞系，选择的能力，以支持复制的温度敏感的VP 5突变体，用于分离VP 5和VP 23无效突变体。VP 5（K5 DELTAZ）和VP 23（K23 Z）中的突变通过在基因编码序列的开始处插入lacZ基因而产生。这两种突变体都不能在非允许细胞系上形成噬斑，因此，VP 23与VP 5一样，是病毒复制所必需的基因产物。这两种突变体表达野生型水平的感染细胞蛋白感染后的许可和nonpermissive细胞系。然而，VP 5（150-kDa）和VP 23（33-kDa）多肽分别在由K5 DELTAZ-和K23 Z-感染的Vero细胞制备的裂解物中不存在。通过对K5 DELTAZ和K23 Z感染的Vero细胞的薄切片进行电子显微镜分析，未观察到衣壳结构。在由突变体感染的细胞的裂解物沉降后，在衣壳通常沉降的级分中未观察到衣壳蛋白。在VP 5和VT 23突变体中以及在KOS感染的Vero细胞中复制的DNA量与在允许细胞中相同。然而，在感染突变体的Vero细胞中，基因组末端不明显，表明DNA保持在多联体中，并且没有加工成单位长度的基因组。
The herpes simplex virus type 1 capsid is composed of seven capsid proteins which are termed VP5, VP19c, VP21, VP22a, VP23, VP24, and VP26. Major capsid protein VP5 is encoded by the gene UL19. UL18, whose transcript is 3' coterminal with that of VP5, specifies capsid protein VP23. Vero cell lines have been isolated that are transformed with either the BglII N (UL19) or EcoRI G (UL16 to UL21) fragment of KOS. These cell lines, selected for the ability to support the replication of a temperature-sensitive VP5 mutant, were used to isolate VP5 and VP23 null mutants. The mutations in VP5 (K5DELTAZ) and VP23 (K23Z) were generated by insertion of the lacZ gene at the beginning of the coding sequences of the genes. Both mutants failed to form plaques on the nonpermissive cell line, and therefore, VP23, like VP5, is an essential gene product for virus replication. Both mutants expressed wild-type levels of infected-cell proteins upon infection of permissive and nonpermissive cell lines. However, the VP5 (150-kDa) and VP23 (33-kDa) polypeptides were absent in lysates prepared from K5DELTAZ- and K23Z-infected Vero cells, respectively. No capsid structures were observed by electron microscopic analysis of thin sections of K5DELTAZ- and K23Z-infected Vero cells. Following sedimentation of lysates from cells infected by the mutants, capsid proteins were not observed in the fractions where capsids normally sediment. The amounts of DNA replicated in the VP5 and VT23 mutant and in KOS-infected Vero cells were the same as in permissive cells. However, genomic ends were not evident in Vero cells infected with the mutants, suggesting that the DNA remains in concatemers and is not processed into unit length genomes.