Simard C, Langlois I, Styger D, Vogt B, Vlcek C, Chalifour A, Trudel M, Schwyzer M
Centre de Recherche en Virologie, Institut Armand-Frappier, Université du Québec, Ville de Laval, Canada.
Virology. 1995 Oct 1;212(2):734-40. doi: 10.1006/viro.1995.1533.
We sequenced the region of the bovine herpesvirus type 1 (BHV-1) genome corresponding to map units 0.172-0.230 (7964 bp), representing the UL39, UL38, and UL37 homologues of herpes simplex virus which encode the large subunit of ribonucleotide reductase (RR) and components of the viral capsid and the tegument, respectively. To discriminate between two potential initiator AUGs of the UL39 gene, the 5' end of the mRNA was mapped by S1 nuclease protection assays. Comparison of the amino acid sequences of the three BHV-1 proteins with analogous polypeptides from several other herpesviruses revealed significant levels of homology. We also compared the expression kinetics of the large (R1, UL39) versus the small (R2, UL40) RR subunits during the course of in vitro BHV-1 infection by Western blotting using specifically developed and calibrated antisera. Our results show that the R1 protein was synthesized earlier than its R2 counterpart. Moreover, the R1 protein accumulated to a higher level than the R2 protein even though the R2 transcript was in greater abundance than the R1 mRNA. This is discussed with regard to the translational efficiency of their transcripts.
我们对牛疱疹病毒1型(BHV - 1)基因组中对应于图谱单位0.172 - 0.230(7964 bp)的区域进行了测序,该区域代表单纯疱疹病毒的UL39、UL38和UL37同源物,分别编码核糖核苷酸还原酶(RR)的大亚基以及病毒衣壳和被膜的成分。为了区分UL39基因的两个潜在起始AUG,通过S1核酸酶保护试验对mRNA的5'端进行了定位。将三种BHV - 1蛋白的氨基酸序列与其他几种疱疹病毒的类似多肽进行比较,发现了显著的同源水平。我们还通过使用专门开发和校准的抗血清进行蛋白质印迹,比较了体外BHV - 1感染过程中大型(R1,UL39)与小型(R2,UL40)RR亚基的表达动力学。我们的结果表明,R1蛋白比其对应的R2蛋白合成得更早。此外,尽管R2转录本比R1 mRNA丰度更高,但R1蛋白积累的水平比R2蛋白更高。针对它们转录本的翻译效率对此进行了讨论。