MOLECULAR-GENETICS OF HERPES-SIMPLEX VIRUS .6. CHARACTERIZATION OF A TEMPERATURE-SENSITIVE MUTANT DEFECTIVE IN THE EXPRESSION OF ALL EARLY VIRAL GENE-PRODUCTS
MOLECULAR-GENETICS OF HERPES-SIMPLEX VIRUS .6. CHARACTERIZATION OF A TEMPERATURE-SENSITIVE MUTANT DEFECTIVE IN THE EXPRESSION OF ALL EARLY VIRAL GENE-PRODUCTS
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DOI：
10.1128/jvi.38.2.539-547.1981
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发表时间：
1981-01-01
影响因子：
5.4
通讯作者：
BUCHAN, A
中科院分区：
文献类型：
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作者：
KNIPE, DM;BATTERSON, W;BUCHAN, A
单纯疱疹病毒1型（HFEM）突变体tsB 7未能表达任何可检测的病毒多肽，并没有显着抑制宿主细胞蛋白质合成的感染细胞[人表皮样癌HEp-2]保持在非允许的温度。该突变体补充了一个共同感染的温度敏感的突变体病毒不同的噬菌斑表型的增长，从而出现能够穿透双重感染的细胞。tsB 7的产量通过共感染病毒而增强，但未达到共感染病毒增强的程度。共感染研究表明，野生型亲本和其他病毒反式补充了tsB 7缺陷，但效果很差。通过用单纯疱疹病毒2型XbaI DNA片段转染的tsB 7的标记拯救定位了0.45-0.70 map U之间的突变。通过该拯救产生的ts+型间重组体的DNA结构分析表明，单纯疱疹病毒2型DNA置换均包含0.46-0.52图U之间的区域，从而进一步确定了突变的图位。对这些型间重组体表达的多肽的分析确定了指定多肽2、6、10、32、43和44的基因的基因组位置，并表明突变定位在编码病毒体结构多肽的基因中或附近。基因组的这一区域仅在病毒DNA复制后以稳定的转录物和细胞质mRNA表示，因此该基因似乎是晚期功能。这些结果与tsB 7中的ts突变是在编码病毒体组分的基因中一致的，所述病毒体组分在α-谷氨酰胺的表达之前起作用。感染早期的基因最可能的解释是突变体在脱壳阶段被阻断，缺陷被共感染病毒基因产物补充，尽管效果不佳。
The herpes simplex virus 1 (HFEM) mutant tsB7 failed to express any detectable viral polpeptides and did not significantly inhibit host cell protein synthesis in infected cells [human epidermoid carcinoma HEp-2] maintained at the nonpermissive temperature. The mutant complemented the growth of a coinfecting temperature-sensitive mutant virus differing in plaque phenotype and thus appeared capable of penetrating doubly infected cells. The yield of tsB7 was enhanced by the coinfecting virus but not to the extent that the coinfecting virus was enhanced. Coinfection studies suggested that the tsB7 defect was complemented in trans, but poorly, by the wild-type parent and other viruses. Marker rescue of tsB7 by transfection with herpes simplex virus 2 XbaI DNA fragments mapped the mutation between 0.45-0.70 map U. Analysis of the DNA structure of the ts+ intertypic recombinants generated by this rescue showed that the herpes simplex virus 2 DNA substitutions all contained the region between 0.46-0.52 map U, thus further defining the map position of the mutation. Analyses of the polypeptides expressed by these intertypic recombinants defined the genome location of the genes specifying polypeptides 2, 6, 10, 32, 43 and 44 and indicated that the mutation maps in or near genes coding for virion structural polypeptides. This region of the genome is represented as stable transcripts and cytoplasmic mRNA only after viral DNA replication and thus this gene appears to be a late function. These results are consistent with the ts mutation in tsB7 being in a gene coding for a virion component which functions before expression of the .alpha. genes early in infection. The most likely explanation is that the mutant is blocked at a stage of uncoating and the defect is complemented, although poorly, by a coinfecting virus gene product.