Department of Hematology, Hebrew University-Hadassah Medical School, Jerusalem, Israel 91120.
J Virol. 2010 Apr;84(7):3431-42. doi: 10.1128/JVI.01735-09. Epub 2010 Jan 20.
The infection process by simian virus 40 (SV40) and entry of its genome into nondividing cells are only partly understood. Infection begins by binding to GM1 receptors at the cell surface, cellular entry via caveolar invaginations, and trafficking to the endoplasmic reticulum, where the virus disassembles. To gain a deeper insight into the contribution of host functions to this process, we studied cellular signaling elicited by the infecting virus. Signaling proteins were detected by Western blotting and immunofluorescence staining. The study was assisted by a preliminary proteomic screen. The contribution of signaling proteins to the infection process was evaluated using specific inhibitors. We found that CV-1 cells respond to SV40 infection by activating poly(ADP-ribose) polymerase 1 (PARP-1)-mediated apoptotic signaling, which is arrested by the Akt-1 survival pathway and stress response. A single key regulator orchestrating the three pathways is phospholipase C-gamma (PLCgamma). The counteracting apoptotic and survival pathways are robustly balanced as the infected cells neither undergo apoptosis nor proliferate. Surprisingly, we have found that the apoptotic pathway, including activation of PARP-1 and caspases, is absolutely required for the infection to proceed. Thus, SV40 hijacks the host defense to promote its infection. Activities of PLCgamma and Akt-1 are also required, and their inhibition abrogates the infection. Notably, this signaling network is activated hours before T antigen is expressed. Experiments with recombinant empty capsids, devoid of DNA, indicated that the major capsid protein VP1 alone triggers this early signaling network. The emerging robust signaling network reflects a delicate evolutionary balance between attack and defense in the host-virus relationship.
猿猴病毒 40(SV40)的感染过程及其基因组进入非分裂细胞的过程尚未完全被理解。感染始于病毒表面 GM1 受体的结合,通过小窝内陷进入细胞,然后转运到内质网,在那里病毒解体。为了更深入地了解宿主功能对这一过程的贡献,我们研究了感染病毒引发的细胞信号。通过 Western blot 和免疫荧光染色检测信号蛋白。该研究得到了初步蛋白质组筛选的辅助。使用特定的抑制剂评估信号蛋白对感染过程的贡献。我们发现,CV-1 细胞通过激活多聚(ADP-核糖)聚合酶 1(PARP-1)介导的凋亡信号来响应 SV40 感染,该信号被 Akt-1 存活途径和应激反应所阻断。一个单一的关键调节因子协调了这三条途径,即磷脂酶 C-γ(PLCγ)。对抗凋亡和存活途径的反应非常强烈,因为感染的细胞既不会凋亡也不会增殖。令人惊讶的是,我们发现凋亡途径,包括 PARP-1 和半胱天冬酶的激活,对于感染的进行是绝对必需的。因此,SV40 劫持了宿主防御来促进其感染。PLCγ 和 Akt-1 的活性也是必需的,它们的抑制会破坏感染。值得注意的是,这个信号网络在 T 抗原表达之前几个小时就被激活了。使用缺乏 DNA 的重组空衣壳的实验表明,仅主要衣壳蛋白 VP1 就可触发这种早期信号网络。新兴的强大信号网络反映了宿主-病毒关系中攻击和防御之间的微妙进化平衡。