Cohen J I, Seidel K E
Medical Virology Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892.
Virology. 1995 Feb 1;206(2):835-42. doi: 10.1006/viro.1995.1006.
Varicella-zoster virus (VZV) encodes 69 unique open reading frames, 5 of which do not have herpes simplex virus type 1 (HSV-1) homologs. One of the 5, ORF1, is predicted to encode a protein of 108 amino acids. We identified a 470-base RNA corresponding to ORF1. To determine whether ORF1 encodes a protein, an 11-amino-acid epitope was inserted in frame after the ninth codon of the ORF1 open reading frame. A recombinant virus carrying this epitope expressed a protein that was immunoprecipitated with monoclonal antibody to the epitope. The ORF1 protein was detected in the membrane of infected cells. The size of ORF1 protein expressed in VZV-infected cells was slightly larger than the size expressed by translation in vitro, suggesting that the protein may undergo post-translational modification in infected cells. Insertion of stop codons immediately before the epitope in the ORF1 gene resulted in a recombinant virus that did not express ORF1 protein and that was not growth impaired in cell culture. Thus, ORF1 encodes a protein that localizes to the membrane of VZV-infected cells and that is dispensable for virus growth in vitro.
水痘带状疱疹病毒(VZV)编码69个独特的开放阅读框,其中5个没有1型单纯疱疹病毒(HSV-1)的同源物。这5个中的一个,即ORF1,预计编码一种108个氨基酸的蛋白质。我们鉴定出了一个与ORF1对应的470个碱基的RNA。为了确定ORF1是否编码一种蛋白质,在ORF1开放阅读框的第九个密码子之后框内插入了一个11个氨基酸的表位。携带该表位的重组病毒表达了一种能被针对该表位的单克隆抗体免疫沉淀的蛋白质。在受感染细胞的膜中检测到了ORF1蛋白。在VZV感染细胞中表达的ORF1蛋白的大小略大于体外翻译所表达的大小,这表明该蛋白可能在受感染细胞中经历翻译后修饰。在ORF1基因的表位之前立即插入终止密码子,产生了一种不表达ORF1蛋白且在细胞培养中生长未受损的重组病毒。因此,ORF1编码一种定位于VZV感染细胞膜的蛋白质,并且该蛋白质对于病毒在体外的生长是可有可无