van Zijl M, van der Gulden H, de Wind N, Gielkens A, Berns A
Division of Molecular Genetics, The Netherlands Cancer Institute, Amsterdam.
J Gen Virol. 1990 Aug;71 ( Pt 8):1747-55. doi: 10.1099/0022-1317-71-8-1747.
We have determined the nucleotide sequence of two genes in the unique short region of the genome of pseudorabies virus (PRV). Near the internal repeat, upstream of the gene encoding glycoprotein gX, we identified an open reading frame (ORF) encoding a protein of 390 amino acids. We designated this gene PK because the predicted protein contains most of the conserved motifs of a eukaryotic protein kinase. The protein shares amino acid homology with the protein kinases encoded by gene US3 of herpes simplex virus type 1 (HSV-1) and gene 66 of varicella-zoster virus. Near the terminal repeat, downstream of a gene encoding an 11K protein, we identified an ORF encoding a protein of 256 amino acids. We designated this gene 28K, the Mr of the predicted protein. Part of the amino acid sequence of 28K is homologous to the predicted US2 protein of HSV-1. Northern blot analysis revealed a 2.7 kb mRNA encoding the putative protein kinase and a 1.2 kb mRNA encoding the 28K protein in PRV-infected cells. The 5' ends of the mRNAs were mapped by primer extension. Two transcriptional start sites were identified for the PK mRNA: a minor start site immediately upstream of the ORF and a major start site (greater than 95% of the mRNA) within the ORF, 64 nucleotides upstream of an internal ATG codon. A single transcriptional start site was identified for the 28K mRNA immediately upstream of the ORF. Immunoblot analysis with anti-peptide sera revealed that, in cells infected with PRV, the PK gene was translated into two proteins with Mrs of 53K and 41K, and the 28K gene into a single protein with an Mr of 28K.
我们已经确定了伪狂犬病病毒（PRV）基因组独特短区域中两个基因的核苷酸序列。在内部重复序列附近，糖蛋白gX编码基因的上游，我们鉴定出一个开放阅读框（ORF），它编码一个由390个氨基酸组成的蛋白质。我们将这个基因命名为PK，因为预测的蛋白质包含真核蛋白激酶的大多数保守基序。该蛋白质与1型单纯疱疹病毒（HSV-1）的US3基因和水痘-带状疱疹病毒的66基因编码的蛋白激酶具有氨基酸同源性。在末端重复序列附近，一个编码11K蛋白的基因下游，我们鉴定出一个ORF，它编码一个由256个氨基酸组成的蛋白质。我们将这个基因命名为28K，即预测蛋白质的相对分子质量。28K的部分氨基酸序列与HSV-1预测的US2蛋白同源。Northern印迹分析显示，在PRV感染的细胞中，有一个2.7 kb的mRNA编码假定的蛋白激酶，还有一个1.2 kb的mRNA编码28K蛋白。通过引物延伸确定了mRNA的5'末端。PK mRNA鉴定出两个转录起始位点：一个次要起始位点在ORF紧邻上游，一个主要起始位点（超过95%的mRNA）在ORF内，位于内部ATG密码子上游64个核苷酸处。28K mRNA在ORF紧邻上游鉴定出一个单一的转录起始位点。用抗肽血清进行的免疫印迹分析表明，在PRV感染的细胞中，PK基因翻译成相对分子质量为53K和41K的两种蛋白质，而28K基因翻译成相对分子质量为28K的单一蛋白质。
