Chang Y E, Menotti L, Filatov F, Campadelli-Fiume G, Roizman B
The Marjorie B. Kovler Viral Oncology Laboratories, The University of Chicago, Chicago, Illinois 60637, USA.
J Virol. 1998 Jul;72(7):6056-64. doi: 10.1128/JVI.72.7.6056-6064.1998.
An antibody made against the herpes simplex virus 1 US5 gene predicted to encode glycoprotein J was found to react strongly with two proteins, one with an apparent Mr of 23,000 and mapping in the S component and one with a herpes simplex virus protein with an apparent Mr of 43,000. The antibody also reacted with herpes simplex virus type 2 proteins forming several bands with apparent Mrs ranging from 43,000 to 50,000. Mapping studies based on intertypic recombinants, analyses of deletion mutants, and ultimately, reaction of the antibody with a chimeric protein expressed by in-frame fusion of the glutathione S-transferase gene to an open reading frame antisense to the gene encoding glycoprotein B led to the definitive identification of the new open reading frame, designated UL27.5. Sequence analyses indicate the conservation of a short amino acid sequence common to US5 and UL27.5. The coding sequence of the herpes simplex virus UL27.5 open reading frame is strongly homologous to the sequence encoding the carboxyl terminus of the herpes simplex virus 2 UL27.5 sequence. However, both open reading frames could encode proteins predicted to be significantly larger than the mature UL27.5 proteins accumulating in the infected cells, indicating that these are either processed posttranslationally or synthesized from alternate, nonmethionine-initiating codons. The UL27.5 gene expression is blocked by phosphonoacetate, indicating that it is a gamma2 gene. The product accumulated predominantly in the cytoplasm. UL27.5 is the third open reading frame found to map totally antisense to another gene and suggests that additional genes mapping antisense to known genes may exist.
一种针对单纯疱疹病毒1型US5基因(预测编码糖蛋白J)制备的抗体,被发现能与两种蛋白质强烈反应,一种表观分子量为23,000,定位于S组分,另一种与一种表观分子量为43,000的单纯疱疹病毒蛋白反应。该抗体还与单纯疱疹病毒2型蛋白反应,形成几条表观分子量范围从43,000到50,000的条带。基于型间重组体的定位研究、缺失突变体分析,以及最终该抗体与通过谷胱甘肽S-转移酶基因与糖蛋白B编码基因的反义开放阅读框进行读框内融合所表达的嵌合蛋白的反应,导致了新开放阅读框的明确鉴定,命名为UL27.5。序列分析表明US5和UL27.5存在一个共同的短氨基酸序列保守区。单纯疱疹病毒UL27.5开放阅读框的编码序列与单纯疱疹病毒2型UL27.5序列羧基末端的编码序列高度同源。然而,这两个开放阅读框都可能编码预测比在感染细胞中积累的成熟UL27.5蛋白大得多的蛋白质,这表明这些蛋白要么是翻译后加工的,要么是从替代的、非甲硫氨酸起始密码子合成的。UL27.5基因的表达被膦乙酸阻断,表明它是一个γ2基因。其产物主要积聚在细胞质中。UL27.5是第三个被发现完全与另一个基因呈反义定位的开放阅读框,这表明可能存在更多与已知基因呈反义定位的基因。