BIO5 Institute, University of Arizona, Tucson, Arizona, United States of America.
Vaccine and Gene Therapy Institute, Oregon Health & Science University, Beaverton, Oregon, United States of America.
PLoS Pathog. 2019 Nov 14;15(11):e1008037. doi: 10.1371/journal.ppat.1008037. eCollection 2019 Nov.
Sustained phosphotinositide3-kinase (PI3K) signaling is critical to the maintenance of alpha and beta herpesvirus latency. We have previously shown that the beta-herpesvirus, human cytomegalovirus (CMV), regulates epidermal growth factor receptor (EGFR), upstream of PI3K, to control states of latency and reactivation. How signaling downstream of EGFR is regulated and how this impacts CMV infection and latency is not fully understood. We demonstrate that CMV downregulates EGFR early in the productive infection, which blunts the activation of EGFR and its downstream pathways in response to stimuli. However, CMV infection sustains basal levels of EGFR and downstream pathway activity in the context of latency in CD34+ hematopoietic progenitor cells (HPCs). Inhibition of MEK/ERK, STAT or PI3K/AKT pathways downstream of EGFR increases viral reactivation from latently infected CD34+ HPCs, defining a role for these pathways in latency. We hypothesized that CMV modulation of EGFR signaling might impact viral transcription important to latency. Indeed, EGF-stimulation increased expression of the UL138 latency gene, but not immediate early or early viral genes, suggesting that EGFR signaling promotes latent gene expression. The early growth response-1 (EGR1) transcription factor is induced downstream of EGFR signaling through the MEK/ERK pathway and is important for the maintenance of hematopoietic stemness. We demonstrate that EGR1 binds the viral genome upstream of UL138 and is sufficient to promote UL138 expression. Further, disruption of EGR1 binding upstream of UL138 prevents the establishment of latency in CD34+ HPCs. Our results indicate a model whereby UL138 modulation of EGFR signaling feeds back to promote UL138 gene expression and suppression of replication for latency. By this mechanism, the virus has hardwired itself into host cell biology to sense and respond to changes in homeostatic host cell signaling.
持续的磷酸肌醇 3-激酶 (PI3K) 信号对于维持α和β疱疹病毒潜伏至关重要。我们之前已经表明,β疱疹病毒,人类巨细胞病毒 (CMV),通过调节表皮生长因子受体 (EGFR),上游的 PI3K,来控制潜伏和再激活状态。EGFR 下游信号如何被调节,以及这如何影响 CMV 感染和潜伏,目前还不完全清楚。我们证明 CMV 在产毒感染的早期下调 EGFR,这削弱了 EGFR 的激活及其下游途径对刺激的反应。然而,CMV 感染在 CD34+造血祖细胞 (HPC) 的潜伏感染背景下维持 EGFR 和下游途径的基础水平活性。EGFR 下游的 MEK/ERK、STAT 或 PI3K/AKT 途径的抑制增加了潜伏感染的 CD34+HPC 中的病毒再激活,这定义了这些途径在潜伏中的作用。我们假设 CMV 对 EGFR 信号的调节可能会影响到潜伏所必需的病毒转录。事实上,EGF 刺激增加了 UL138 潜伏基因的表达,但不增加即刻早期或早期病毒基因的表达,这表明 EGFR 信号促进潜伏基因的表达。早期生长反应-1 (EGR1) 转录因子通过 MEK/ERK 途径在 EGFR 信号下游诱导,对维持造血干细胞特性很重要。我们证明 EGR1 在 UL138 的上游结合病毒基因组,足以促进 UL138 的表达。此外,破坏 UL138 上游的 EGR1 结合阻止了 CD34+HPC 中潜伏的建立。我们的结果表明了一个模型,即 UL138 对 EGFR 信号的调节反馈促进 UL138 基因表达和抑制潜伏时的复制。通过这种机制,病毒已经将自身硬连接到宿主细胞生物学中,以感知和响应宿主细胞信号的稳态变化。