Department of Biopharmaceutical Convergence, Sungkyunkwan University, Suwon, South Korea.
College of Biotechnology and Bioengineering, Sungkyunkwan University, Suwon, South Korea.
PLoS Pathog. 2022 Dec 1;18(12):e1011007. doi: 10.1371/journal.ppat.1011007. eCollection 2022 Dec.
YES-associated protein (YAP), a critical actor of the mammalian Hippo signaling pathway involved in diverse biological events, has gained increased recognition as a cellular factor regulated by viral infections, but very few studies have investigated their relationship vice versa. In this study, we show that YAP impairs HCMV replication as assessed by viral gene expression analysis and progeny assays, and that this inhibition occurs at the immediate-early stages of the viral life cycle, at the latest. Using YAP mutants lacking key functional domains and shRNA against TEAD, we show that the inhibitory effects of YAP on HCMV replication are nuclear localization- and TEAD cofactor-dependent. Quantitative real-time PCR (qPCR) and subcellular fractionation analyses reveal that YAP does not interfere with the viral entry process but inhibits transport of the HCMV genome into the nucleus. Most importantly, we show that the expression of stimulator of interferon genes (STING), recently identified as an important component for nuclear delivery of the herpesvirus genome, is severely downregulated by YAP at the level of gene transcription. The functional importance of STING is further confirmed by the observation that STING expression restores YAP-attenuated nuclear transport of the HCMV genome, viral gene expression, and progeny virus production. We also show that HCMV-upregulated YAP reduces expression of STING. Taken together, these findings indicate that YAP possesses both direct and indirect regulatory roles in HCMV replication at different infection stages.
Yes 相关蛋白 (YAP) 是哺乳动物 Hippo 信号通路的关键因子,参与多种生物事件,作为受病毒感染调控的细胞因子受到越来越多的关注,但很少有研究调查它们之间的相互关系。在这项研究中,我们表明 YAP 可通过病毒基因表达分析和后代测定来损害 HCMV 的复制,并且这种抑制作用最迟发生在病毒生命周期的早期。使用缺乏关键功能结构域的 YAP 突变体和针对 TEAD 的 shRNA,我们表明 YAP 对 HCMV 复制的抑制作用依赖于核定位和 TEAD 辅因子。实时定量 PCR (qPCR) 和亚细胞分级分析表明 YAP 不干扰病毒进入过程,但抑制 HCMV 基因组向核内的转运。最重要的是,我们表明干扰素基因刺激物 (STING) 的表达被 YAP 在基因转录水平严重下调,STING 最近被确定为疱疹病毒基因组核内传递的重要组成部分。STING 的功能重要性通过观察到 STING 表达恢复 YAP 减弱的 HCMV 基因组核内转运、病毒基因表达和子代病毒产生得到进一步证实。我们还表明,HCMV 上调的 YAP 降低了 STING 的表达。总之,这些发现表明 YAP 在不同感染阶段对 HCMV 复制具有直接和间接的调节作用。