Breaking human cytomegalovirus major immediate-early gene silence by vasoactive intestinal peptide stimulation of the protein kinase A-CREB-TORC2 signaling cascade in human pluripotent embryonal NTera2 cells.
Breaking human cytomegalovirus major immediate-early gene silence by vasoactive intestinal peptide stimulation of the protein kinase A-CREB-TORC2 signaling cascade in human pluripotent embryonal NTera2 cells.
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通过血管活性肠肽刺激人多能胚胎 NTera2 细胞中的蛋白激酶 A-CREB-TORC2 信号级联，打破人巨细胞病毒主要立即早期基因沉默。
DOI：
10.1128/jvi.00061-09
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发表时间：
2009
影响因子：
5.4
通讯作者：
Meier,JefferyL
中科院分区：
文献类型：
--
作者：
Yuan,Jinxiang;Liu,Xiaoqiu;Wu,AllenW;McGonagill,PatrickW;Keller,MichaelJ;Galle,CourtneyS;Meier,JefferyL
人类巨细胞病毒(HCMV)在潜伏感染者中重新激活的触发机制尚不清楚。在潜伏期间，HCMV主要即刻早期(MIE)基因的表达打破沉默，启动病毒的重新激活。用静止感染的人多能胚胎NTera2细胞(NT2)复制HCMV再激活模型，我们发现免疫调节神经肽血管活性肠肽(VIP)可立即激活HCMV mie基因的表达，并呈剂量依赖关系(1~500 nM)。这种反应需要MIE增强子环状AMP反应元件(Cre)。VIP可迅速提高CREB Ser133和ATF-1 Ser63的磷酸化水平，尽管CREB Ser133的磷酸化水平在基线水平相当可观。VIP不改变细胞核、Oct4(多能细胞标志物)或hDaxx(HCMV基因表达的细胞抑制因子)中的HCMV基因组水平。VIP激活的MIE基因的表达由细胞蛋白激酶A(PKA)、CREB和TORC2介导。VIP诱导依赖PKA的TORC2 Ser171去磷酸化和核进入，这可能使MIE基因激活，因为TORC2 S171a(没有Ser171磷酸化)在没有VIP刺激的情况下表现出增强的核进入并使MIE基因沉默。总之，VIP刺激PKA-CREB-TORC2信号级联激活静止期感染的NT2细胞中依赖于HCMV CRE的MIE基因的表达。我们推测，通过这种信号级联的神经激素刺激是逆转体内HCMV沉默的一种可能的方法。
The triggering mechanisms underlying reactivation of human cytomegalovirus (HCMV) in latently infected persons are unclear. During latency, HCMV major immediate-early (MIE) gene expression breaks silence to initiate viral reactivation. Using quiescently HCMV-infected human pluripotent embryonal NTera2 cells (NT2) to model HCMV reactivation, we show that vasoactive intestinal peptide (VIP), an immunomodulatory neuropeptide, immediately and dose-dependently (1 to 500 nM) activates HCMV MIE gene expression. This response requires the MIE enhancer cyclic AMP response elements (CRE). VIP quickly elevates CREB Ser133 and ATF-1 Ser63 phosphorylation levels, although the CREB Ser133 phosphorylation level is substantial at baseline. VIP does not change the level of HCMV genomes in nuclei, Oct4 (pluripotent cell marker), or hDaxx (cellular repressor of HCMV gene expression). VIP-activated MIE gene expression is mediated by cellular protein kinase A (PKA), CREB, and TORC2. VIP induces PKA-dependent TORC2 Ser171 dephosphorylation and nuclear entry, which likely enables MIE gene activation, as TORC2 S171A (devoid of Ser171 phosphorylation) exhibits enhanced nuclear entry and desilences the MIE genes in the absence of VIP stimulation. In conclusion, VIP stimulation of the PKA-CREB-TORC2 signaling cascade activates HCMV CRE-dependent MIE gene expression in quiescently infected NT2 cells. We speculate that neurohormonal stimulation via this signaling cascade is a possible means for reversing HCMV silence in vivo.