Foster T P, Chouljenko V N, Kousoulas K G
School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana, 70803, USA.
Virology. 1999 Jun 5;258(2):365-74. doi: 10.1006/viro.1999.9743.
We cloned the gene specified by African monkey kidney cells (Vero) that codes for the homolog of the herpes virus entry mediator (HveA) specified by HeLa cells. The primary sequence of the monkey HveA (HveAs) differed significantly from HveA. Single amino acid differences were distributed throughout the amino and carboxyl terminal portions of the HveAs in comparison with the HveA, whereas certain regions were highly conserved. The predicted membrane spanning domains of the two receptors differed substantially due to insertions and deletions of short amino acid sequences. The ability of HveAs to mediate HSV virus entry was tested in a series of experiments using the recombinant virus KOS/EGFP, which constitutively expressed the enhanced green fluorescence protein (EGFP) and Chinese hamster ovary cells (CHO) transformed with the HveAs gene. The KOS/EGFP virus was constructed by inserting an EGFP gene cassette within the intergenic region between the UL53 (gK) and UL54 (ICP27) genes. The KOS/EGFP virus formed viral plaques and replicated as well as the wild-type KOS virus. HveAs-transformed CHO cells constitutively expressing HveAs mediated herpesvirus entry efficiently, whereas cells transformed with the HveAs gene in the noncoding orientation did not mediate virus entry. A genetically engineered protein composed of the amino-terminal portion of the HveAs protein fused to the heavy chain of mouse IgG immunoglobulin as well as mouse antibodies raised against HveAs blocked virus entry into HveAs-transformed CHO cells. Thus, HveAs is the functional homolog of HveA.
我们克隆了非洲猴肾细胞（Vero）中编码的基因，该基因对应于HeLa细胞中编码的疱疹病毒进入介质（HveA）的同源物。猴HveA（HveAs）的一级序列与HveA有显著差异。与HveA相比，单个氨基酸差异分布在HveAs的氨基和羧基末端部分，而某些区域高度保守。由于短氨基酸序列的插入和缺失，两种受体的预测跨膜结构域有很大不同。在一系列实验中，使用重组病毒KOS/EGFP（其组成性表达增强型绿色荧光蛋白（EGFP））和用HveAs基因转化的中国仓鼠卵巢细胞（CHO），测试了HveAs介导单纯疱疹病毒进入的能力。KOS/EGFP病毒是通过在UL53（gK）和UL54（ICP27）基因之间的基因间隔区内插入一个EGFP基因盒构建而成。KOS/EGFP病毒形成病毒斑并能像野生型KOS病毒一样复制。组成性表达HveAs的经HveAs转化的CHO细胞能有效地介导疱疹病毒进入，而用非编码方向的HveAs基因转化的细胞则不能介导病毒进入。一种由HveAs蛋白的氨基末端部分与小鼠IgG免疫球蛋白重链融合而成的基因工程蛋白，以及针对HveAs产生的小鼠抗体，可阻断病毒进入经HveAs转化的CHO细胞。因此，HveAs是HveA的功能同源物。
