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病毒刺激相关蛋白HSRG1是HSV-1病毒感染细胞时诱导表达的蛋白之一,本研究组曾报道其具有与转录调控蛋白相互作用的特性.为进一步分析HSRG1对病毒基因转录调控的影响,本研究在转染细胞中上调HSRG1分子的表达,发现它能够延缓HSV-1病毒的增殖.CAT分析结果也表明HSRG1对HSV-1多种病毒启动子的转录效率具有量效抑制作用.运用酵母双杂交及免疫沉淀技术表明,HSRG1与对RNA聚合酶Ⅱ(RNAPⅡ)的转录延伸具有重要作用的复合物P-TEFb(positive transcription elongation factor b,P-TEFb)中的调节亚基细胞周期蛋白T2(Cyclin T2)存在蛋白质相互作用,且Cyclin T2氨基端420氨基酸对二者的结合具有重要作用.荧光共定位实验表明,共转染的细胞中两蛋白的细胞内定位因二者相互作用而受影响.进一步利用CAT实验证实,HSRG1能够拮抗Cyclin T2对多种病毒启动子的转录激活作用,提示HSRG1对病毒复制增殖的抑制作用很可能是通过与Cyclin T2的结合而产生的,它具有阻断Cyclin T2的调控,影响病毒基因转录的延伸.
The virus-stimulated protein HSRG1 is one of the proteins induced by HSV-1 virus infection and the research group has reported its interaction with transcriptional regulatory proteins.In order to further analyze the effect of HSRG1 on the transcriptional regulation of the viral gene, The results showed that HSRG1 could delay the proliferation of HSV-1 virus.The results of CAT assay also showed that HSRG1 could inhibit the transcriptional efficiency of HSV-1 promoters in a dose-dependent manner.Using yeast two-hybrid And immunoprecipitation techniques showed that the regulatory subunit cyclin T2 (Cyclin) in HSC1 and P-TEFb (P-TEFb), which plays an important role in the transcription elongation of RNA polymerase Ⅱ (RNAPⅡ) T2) and there is an important role for the combination of the two amino acids at the amino terminus of Cyclin T2.Fluorescent co-localization experiments show that the intracellular localization of the two proteins in the cotransfected cells is affected by the interaction between the two. Further use of CAT experiments confirmed that HSRG1 can antagonize the transcriptional activation of Cyclin T2 on a variety of viral promoters, suggesting that HSRG1 inhibition of viral replication It is likely that this effect is caused by binding to Cyclin T2, which has the effect of blocking the regulation of Cyclin T2 and affecting the transcription of the viral gene.