Adenovirus targets transcriptional and posttranslational mechanisms to limit gap junction function.
Adenovirus targets transcriptional and posttranslational mechanisms to limit gap junction function.
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DOI：
10.1096/fj.202000667r
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发表时间：
2020-07
期刊：
影响因子：
--
通讯作者：
Smyth JW
中科院分区：
文献类型：
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作者：
Calhoun PJ;Phan AV;Taylor JD;James CC;Padget RL;Zeitz MJ;Smyth JW
腺病毒是包括病毒性心肌炎在内的一系列致病因素。缝隙连接蛋白Cx43(Cx43，基因名GJA1)促进了每一次心跳所必需的动作电位的快速传播。缝隙连接也传播先天和获得性抗病毒免疫反应，但病毒如何针对这些结构尚不清楚。鉴于Cx43的这种免疫学作用，我们假设缝隙连接将在5型腺病毒(Ad5)感染过程中成为靶点。我们发现，Cx43蛋白水平的降低是由于在Ad5感染期间依赖于β-连环蛋白转录活性的GJA1基因转录减少，早期病毒蛋白E4orf1足以诱导β-连环蛋白磷酸化。间隙连接功能的丧失发生在Cx43蛋白水平降低之前，Ad5感染会迅速诱导Cx43磷酸化事件，这与间隙连接电导的变化一致。Cx43与ZO-1的直接相互作用在缝隙连接调控中起着关键作用。我们通过免疫共沉淀和互补研究发现，在Ad5感染过程中，Cx43/ZO-1复合体丢失，在人诱导的多能干细胞来源的心肌细胞中，通过减少ZO-1复合体，Cx43缝隙连接重塑。这些发现揭示了Ad5对缝隙连接功能的特异性靶向，导致细胞间通讯的丧失，这可能导致感染心脏的危险病理状态，包括心律失常。
Adenoviruses are responsible for a spectrum of pathogenesis including viral myocarditis. The gap junction protein connexin43 (Cx43, gene name GJA1) facilitates rapid propagation of action potentials necessary for each heartbeat. Gap junctions also propagate innate and adaptive antiviral immune responses, but how viruses may target these structures is not understood. Given this immunological role of Cx43, we hypothesized that gap junctions would be targeted during adenovirus type 5 (Ad5) infection. We find reduced Cx43 protein levels due to decreased GJA1 mRNA transcripts dependent upon β-catenin transcriptional activity during Ad5 infection, with early viral protein E4orf1 sufficient to induce β-catenin phosphorylation. Loss of gap junction function occurs prior to reduced Cx43 protein levels with Ad5 infection rapidly inducing Cx43 phosphorylation events consistent with altered gap junction conductance. Direct Cx43 interaction with ZO-1 plays a critical role in gap junction regulation. We find loss of Cx43/ZO-1 complexing during Ad5 infection by co-immunoprecipitation and complementary studies in human induced pluripotent stem cell derived-cardiomyocytes reveal Cx43 gap junction remodeling by reduced ZO-1 complexing. These findings reveal specific targeting of gap junction function by Ad5 leading to loss of intercellular communication which would contribute to dangerous pathological states including arrhythmias in infected hearts.