Sampath Padmavathi, Deluca Neal A
E1257 Biomedical Science Tower, Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
J Virol. 2008 Mar;82(5):2339-49. doi: 10.1128/JVI.02459-07. Epub 2007 Dec 19.
The binding of herpes simplex virus type 1 ICP4, TATA-binding protein (TBP), and RNA polymerase II (polII) to the promoter regions of representative immediate-early (IE) (ICP0), early (E) (thymidine kinase [tk]), and late (L) (glycoprotein C [gC]) genes on the viral genome was examined as a function of time postinfection, viral DNA replication, cis-acting sites for TFIID in the tk and gC promoters, and genetic background of ICP4. The binding of TBP and polII to the IE ICP0 promoter was independent of the presence of ICP4, whereas the binding of TBP and polII to the tk and gC promoters occurred only when ICP4 also bound to the promoters, suggesting that the presence of ICP4 at the promoters of E and L genes in virus-infected cells is crucial for the formation of transcription complexes on these promoters. When the TATA box of the tk promoter or the initiator element (INR) of the gC promoter was mutated, a reduction in the amount of TBP and polII binding was observed. However, a reduction in the amount of ICP4 binding to the promoters was also observed, suggesting that the binding of TBP-containing complexes and ICP4 is cooperative. The binding of ICP4, TBP, and polII was also observed on the gC promoter at early times postinfection or when DNA synthesis was inhibited, suggesting that transcription complexes may be formed early on L promoters and that additional events or proteins are required for expression. The ability to form these early complexes on the gC promoter required the DNA-binding domain but in addition required the carboxyl-terminal 524 amino acids of ICP4, which is missing the virus n208. This region was not required to form TBP- and polII-containing complexes on the tk promoter. n208 activates E but not L genes during viral infection. These data suggest that a region of ICP4 may differentiate between forming TBP- and polII-containing complexes on E and L promoters.
研究了单纯疱疹病毒1型ICP4、TATA结合蛋白(TBP)和RNA聚合酶II(polII)与病毒基因组上代表性的立即早期(IE)(ICP0)、早期(E)(胸苷激酶[tk])和晚期(L)(糖蛋白C[gC])基因启动子区域的结合情况,该结合情况是感染后时间、病毒DNA复制、tk和gC启动子中TFIID的顺式作用位点以及ICP4遗传背景的函数。TBP和polII与IE ICP0启动子的结合不依赖于ICP4的存在,而TBP和polII与tk和gC启动子的结合仅在ICP4也与启动子结合时发生,这表明在病毒感染细胞中,E和L基因启动子处ICP4的存在对于这些启动子上转录复合物的形成至关重要。当tk启动子的TATA框或gC启动子的起始元件(INR)发生突变时,观察到TBP和polII结合量减少。然而,也观察到ICP4与启动子结合量减少,这表明含TBP的复合物与ICP4的结合是协同的。在感染后早期或DNA合成受到抑制时,也观察到ICP4、TBP和polII在gC启动子上的结合,这表明转录复合物可能在L启动子上早期形成,并且表达还需要其他事件或蛋白质。在gC启动子上形成这些早期复合物的能力需要DNA结合结构域,但此外还需要ICP4的羧基末端524个氨基酸,病毒n208缺失该区域。该区域对于在tk启动子上形成含TBP和polII的复合物不是必需的。n208在病毒感染期间激活E基因但不激活L基因。这些数据表明,ICP4的一个区域可能在E和L启动子上形成含TBP和polII的复合物方面存在差异。