Zhao Y, Holden V R, Smith R H, O'Callaghan D J
Department of Microbiology and Immunology, Louisiana State University Medical Center, Shreveport 71130-3932, USA.
J Virol. 1995 May;69(5):2786-93. doi: 10.1128/JVI.69.5.2786-2793.1995.
The UL3 protein of equine herpesvirus 1 (EHV-1) KyA strain is a homolog of the ICP27 alpha regulatory protein of herpes simplex virus type 1 (HSV-1) and the ORF 4 protein of varicella-zoster virus. To characterize the regulatory function of the UL3 gene product, a UL3 gene expression vector (pSVUL3) and a vector expressing a truncated version of the UL3 gene (pSVUL3P) were generated. These effector plasmids, in combination with an EHV-1 immediate-early (IE) gene expression vector (pSVIE) and chimeric EHV-1 promoter-chloramphenicol acetyltransferase (CAT) reporter constructs, were used in transient transfection assays. These assays demonstrated that the EHV-1 UL3 gene product is a regulatory protein that can independently trans activate the EHV-1 IE promoter; however, this effect can be inhibited by the repressive function of the IE gene product on the IE promoter (R. H. Smith, G. B. Caughman, and D. J. O'Callaghan, J. Virol. 66:936-945, 1992). In the presence of the IE gene product, the UL3 gene product significantly augments gene expression directed by the promoters of three EHV-1 early genes (thymidine kinase; IR4, which is the homolog of HSV-1 ICP22; and UL3 [ICP27]) and the promoter of the EHV-1 late gene IR5, which is the homolog of HSV-1 US10. Sequences located at nucleotides -123 to +20 of the UL3 promoter harbor a TATA box, SP1 binding site, CAAT box, and octamer binding site and, when linked to the CAT reporter gene, are trans activated to maximal levels by the pSVIE construct in transient expression assays. Results from CAT assays also suggest that the first 11 amino acids of the UL3 protein are not essential for the regulatory function of the UL3 gene product.
马疱疹病毒1型(EHV-1)KyA株的UL3蛋白是单纯疱疹病毒1型(HSV-1)的ICP27α调节蛋白和水痘-带状疱疹病毒的ORF 4蛋白的同源物。为了表征UL3基因产物的调节功能,构建了一个UL3基因表达载体(pSVUL3)和一个表达UL3基因截短版本的载体(pSVUL3P)。这些效应质粒与一个EHV-1立即早期(IE)基因表达载体(pSVIE)以及嵌合的EHV-1启动子-氯霉素乙酰转移酶(CAT)报告基因构建体一起用于瞬时转染试验。这些试验表明,EHV-1 UL3基因产物是一种调节蛋白,能够独立反式激活EHV-1 IE启动子;然而,这种效应可被IE基因产物对IE启动子的抑制功能所抑制(R. H. 史密斯、G. B. 考曼和D. J. 奥卡拉汉,《病毒学杂志》66:936 - 945,1992年)。在存在IE基因产物的情况下,UL3基因产物显著增强由三个EHV-1早期基因(胸苷激酶;IR4,它是HSV-1 ICP22的同源物;以及UL3 [ICP27])的启动子和EHV-1晚期基因IR5(它是HSV-1 US10的同源物)的启动子所指导的基因表达。位于UL3启动子核苷酸-123至+20处的序列含有一个TATA框、SP1结合位点、CAAT框和八聚体结合位点,并且当与CAT报告基因连接时,在瞬时表达试验中被pSVIE构建体反式激活至最大水平。CAT试验结果还表明,UL3蛋白的前11个氨基酸对于UL3基因产物的调节功能并非必需。