Poma E E, Kowalik T F, Zhu L, Sinclair J H, Huang E S
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, 27599-7295, USA.
J Virol. 1996 Nov;70(11):7867-77. doi: 10.1128/JVI.70.11.7867-7877.1996.
The Rb-related p107 protein has been implicated as an important control element in proper cell cycle progression. The p107 protein is thought to restrict cellular proliferation in part through its interaction with the E2F family of transcription factors and is, therefore, a specific target for regulation by several DNA viruses. Here, we demonstrate that p107 protein levels are induced in a biphasic manner in human fibroblasts during productive infection by the human cytomegalovirus (HCMV). Expression patterns of p107 protein levels during HCMV infection of human embryonic lung cells (HELs) demonstrate a sustained induction from early to late times of infection. We also demonstrate that the HCMV immediate-early protein IE1-72 complexes in vivo with the p107 protein and that this interaction can be reconstituted in an in vitro system by using reticulocyte-translated protein. Our data demonstrate that the interaction between p107 and the IE1-72 protein occurs at times of infection that temporally match the second tier of p107 protein induction and the phosphorylation pattern of the IE1-72 protein. Furthermore, we show here that the ability of p107 to transcriptionally repress E2F-responsive promoters can be overcome by expression of the IE1-72 protein. This effect appears to be specific, since the IE1-72 protein is not capable of relieving Rb-mediated repression of an E2F-responsive promoter. Finally, our data demonstrate that HCMV infection can induce cellular proliferation in quiescent cells and that IE1-72 expression alone can, to a degree, drive a similar progression through the cell cycle. These data suggest that IE1-72-mediated transactivation of E2F-responsive promoters through alleviation of p107 transcriptional repression may play a key role in the cell cycle progression stimulated by HCMV infection.
与Rb相关的p107蛋白被认为是细胞周期正常进展中的一个重要调控元件。p107蛋白被认为部分通过与E2F转录因子家族的相互作用来限制细胞增殖,因此是几种DNA病毒调控的特定靶点。在此,我们证明在人巨细胞病毒(HCMV)的生产性感染过程中,人成纤维细胞中的p107蛋白水平以双相方式被诱导。人胚肺细胞(HELs)在HCMV感染期间p107蛋白水平的表达模式显示,从感染早期到晚期持续诱导。我们还证明HCMV立即早期蛋白IE1-72在体内与p107蛋白形成复合物,并且这种相互作用可以在体外系统中通过使用网织红细胞翻译的蛋白来重建。我们的数据表明,p107与IE1-72蛋白之间的相互作用发生在感染时期,这在时间上与p107蛋白诱导的第二阶段以及IE1-72蛋白的磷酸化模式相匹配。此外,我们在此表明,IE1-72蛋白的表达可以克服p107转录抑制E2F反应性启动子的能力。这种效应似乎是特异性的,因为IE1-72蛋白不能解除Rb介导的对E2F反应性启动子的抑制。最后,我们的数据表明HCMV感染可以诱导静止细胞中的细胞增殖,并且单独的IE1-72表达在一定程度上可以驱动类似的细胞周期进程。这些数据表明,IE1-72通过减轻p107转录抑制对E2F反应性启动子的反式激活可能在HCMV感染刺激的细胞周期进程中起关键作用。