Frink R J, Eisenberg R, Cohen G, Wagner E K
J Virol. 1983 Feb;45(2):634-47. doi: 10.1128/JVI.45.2.634-647.1983.
We previously showed that the right third of HindIII fragment L (0.59 to 0.65) of herpes simplex virus type 1 (HSV-1) encodes a family of mRNAs some members of which appear to be related by splicing. In the experiments described in this communication, we determined the nucleotide sequence of the DNA encoding this mRNA family and precisely located the mRNAs associated with this DNA sequence. The major mRNA species is unspliced and encoded by a 2.520-nucleotide region. Just upstream of the 5' end are TATA and CAT box sequences characteristic of HSV-1 promoters. The 3' end maps near a region containing a nominal polyadenylation signal. Three minor species (2,400, 2,200, and 1,900 bases, respectively) appear to share a very short leader sequence with the 5' end of the major mRNA and are then encoded by uninterrupted DNA sequences beginning about 100, 400, and 625 bases downstream of the 5' end of the major unspliced mRNA. These positions map at or very near positions which agree reasonably well with consensus splice acceptor sequences. The fourth mRNA is encoded by a contiguous 730-nucleotide sequence at the 3' end of the major unspliced mRNA and has its 5' end just downstream of recognizable TATA and CAT box sequences. We suggest that this mRNA is controlled by its own promoter. The nucleotide sequence data, in combination with the mRNA localization, demonstrate four potential polypeptides encoded by the region. The largest is 1,569 bases long and defines a 523-amino acid protein with sequence features characteristic of a glycoprotein. This was confirmed to be HSV-1 glycoprotein C by immune precipitation of the in vitro translation product of the major unspliced mRNA, performed with a polyspecific antibody to HSV-1 envelope glycoproteins (anti-env-1 serum), and by comparison of tryptic peptides of this translation product with those of authentic HSV-1 glycoprotein C. Polypeptides encoded by some of the minor species also were tentatively identified.
我们先前表明，单纯疱疹病毒1型（HSV-1）的HindIII片段L的右三分之一（0.59至0.65）编码一族mRNA，其中一些成员似乎通过剪接相关联。在本通讯所述的实验中，我们确定了编码该mRNA家族的DNA的核苷酸序列，并精确地定位了与该DNA序列相关的mRNA。主要的mRNA种类是未剪接的，由一个2520个核苷酸的区域编码。在5'端上游是HSV-1启动子特有的TATA和CAT框序列。3'端定位在一个含有标称聚腺苷酸化信号的区域附近。三种较小的种类（分别为2400、2200和1900个碱基）似乎与主要mRNA的5'端共享一个非常短的前导序列，然后由主要未剪接mRNA的5'端下游约100、400和625个碱基处开始的不间断DNA序列编码。这些位置定位在或非常接近与共有剪接受体序列相当吻合的位置。第四种mRNA由主要未剪接mRNA 3'端的一个连续730个核苷酸的序列编码，其5'端恰好在可识别的TATA和CAT框序列的下游。我们认为这种mRNA受其自身启动子的控制。核苷酸序列数据与mRNA定位相结合，证明该区域编码四种潜在的多肽。最大的多肽长1569个碱基，定义了一个523个氨基酸的蛋白质，具有糖蛋白的序列特征。通过用针对HSV-1包膜糖蛋白的多特异性抗体（抗env-1血清）对主要未剪接mRNA的体外翻译产物进行免疫沉淀，以及将该翻译产物的胰蛋白酶肽段与真实的HSV-1糖蛋白C的肽段进行比较，证实这是HSV-1糖蛋白C。一些较小种类编码的多肽也被初步鉴定。
