Khalil Mohamed I, Ruyechan William T, Hay John, Arvin Ann
Departments of Pediatrics and Microbiology & Immunology, Stanford University School of Medicine, Stanford, CA, United States; Department of Molecular Biology, National Research Center EL-Buhouth St., Dokki, Cairo, Egypt.
Department of Microbiology and Immunology and the Witebsky Center for Microbial Pathogenesis and Immunology, University at Buffalo, Buffalo, NY, United States.
Virology. 2015 Nov;485:47-57. doi: 10.1016/j.virol.2015.06.031. Epub 2015 Jul 23.
The immediate early (IE) 62 protein is the major varicella-zoster virus (VZV) regulatory factor. Analysis of the VZV genome revealed 40 predicted GC-rich boxes within 36 promoters. We examined effects of ectopic expression of Sp1-Sp4 on IE62- mediated transactivation of three viral promoters. Ectopic expression of Sp3 and Sp4 enhanced IE62 activation of ORF3 and gI promoters while Sp3 reduced IE62 activation of ORF28/29 promoter and VZV DNA replication. Sp2 reduced IE62 transactivation of gI while Sp1 had no significant influence on IE62 activation with any of these viral promoters. Electrophoretic mobility shift assays (EMSA) confirmed binding of Sp1 and Sp3 but not Sp2 and Sp4 to the gI promoter. Sp1-4 bound to IE62 and amino acids 238-258 of IE62 were important for the interaction with Sp3 and Sp4 as well as Sp1. This work shows that Sp family members have differential effects on IE62-mediated transactivation in a promoter-dependent manner.
即刻早期(IE)62蛋白是水痘-带状疱疹病毒(VZV)的主要调节因子。对VZV基因组的分析显示,在36个启动子中有40个预测的富含GC的框。我们研究了Sp1-Sp4异位表达对IE62介导的三个病毒启动子反式激活的影响。Sp3和Sp4的异位表达增强了IE62对ORF3和gI启动子的激活,而Sp3降低了IE62对ORF28/29启动子的激活以及VZV DNA复制。Sp2降低了IE62对gI的反式激活,而Sp1对这些病毒启动子中任何一个的IE62激活均无显著影响。电泳迁移率变动分析(EMSA)证实Sp1和Sp3与gI启动子结合,但Sp2和Sp4不结合。Sp1-4与IE62结合,IE62的238-258位氨基酸对于与Sp3、Sp4以及Sp1的相互作用很重要。这项工作表明,Sp家族成员以启动子依赖的方式对IE62介导的反式激活具有不同的影响。