Characterization of the Herpes Simplex Virus (HSV) Tegument Proteins That Bind to gE/gI and US9, Which Promote Assembly of HSV and Transport into Neuronal Axons
Characterization of the Herpes Simplex Virus (HSV) Tegument Proteins That Bind to gE/gI and US9, Which Promote Assembly of HSV and Transport into Neuronal Axons
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DOI：
10.1128/jvi.01113-20
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发表时间：
2020-12-01
影响因子：
5.4
通讯作者：
Johnson, David C.
中科院分区：
文献类型：
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作者：
DuRaine, Grayson;Wisner, Todd W.;Johnson, David C.
单纯疱疹病毒(HSV)异源二聚体Ge/Gi和另一种具有神经元特异性作用的膜蛋白US9促进病毒颗粒在神经元轴突中的顺行运输。HSV gE和US9的缺失阻止了被包裹的颗粒在神经元细胞质中的组装，这解释了为什么HSV病毒粒子不进入轴突。细胞质被膜依赖于病毒膜蛋白和包裹衣壳的被蛋白之间的相互作用。我们报道了被Ge-/US9双重突变体感染的神经元中被膜蛋白UL16是不稳定的，即迅速降解的。用HSV感染神经元的裂解物进行的免疫沉淀实验表明，UL16和另外三种被蛋白，即VP22，UL11和UL21，可以与GE或GI结合。所有这四种被膜蛋白也都被US9提取了下来。在被膜蛋白和GE/GI或US9的神经元中，有充分的证据表明VP22和UL16直接与US9和GE/GI结合。然而，与Ge/GI和US9共沉淀的这些被膜蛋白的数量比感染细胞的少。这显然与感染细胞中形成的几种不同被膜蛋白的基质有关，这些被膜蛋白与GE/GI和US9结合。在转染单种被膜蛋白的细胞中，这种基质不太常见。同样，在HSV感染的细胞中观察到Ge/Gi和US9的共沉淀，而在转染组细胞中没有观察到，这与US9-Ge/Gi的直接相互作用相反。这些研究表明，Ge/Gi和US9与这些被膜蛋白的结合对HSV组装具有神经元特异性影响，HSV组装是被膜颗粒轴突运输所需的过程。这些病毒的一个基本特性是能够在感觉神经元中建立潜伏期，并从潜伏期重新激活，然后在皮肤和粘膜上皮等周围组织中引起疾病。新生的HSV颗粒从神经元细胞体进入轴突，并沿着轴突运输到外围的轴突尖端，是这种重新激活和再感染的重要组成部分。两种HSV膜蛋白Ge/Gi和US9在这些过程中起着至关重要的作用。我们的研究有助于阐明HSV、GE/GI和US9是如何促进病毒颗粒的组装和这些病毒粒子分选成神经元轴突的。
The herpes simplex virus (HSV) heterodimer gE/gI and another membrane protein, US9, which has neuron-specific effects, promote the anterograde transport of virus particles in neuronal axons. Deletion of both HSV gE and US9 blocks the assembly of enveloped particles in the neuronal cytoplasm, which explains why HSV virions do not enter axons. Cytoplasmic envelopment depends upon interactions between viral membrane proteins and tegument proteins that encrust capsids. We report that tegument protein UL16 is unstable, i.e., rapidly degraded, in neurons infected with a gE-/US9double mutant. Immunoprecipitation experiments with lysates of HSV-infected neurons showed that UL16 and three other tegument proteins, namely, VP22, UL11, and UL21, bound either to gE or gI. All four of these tegument proteins were also pulled down with US9. In neurons transfected with tegument proteins and gE/gI or US9, there was good evidence that VP22 and UL16 bound directly to US9 and gE/gI. However, there were lower quantities of these tegument proteins that coprecipitated with gE/gI and US9 from transfected cells than those of infected cells. This apparently relates to a matrix of several different tegument proteins formed in infected cells that bind to gE/gI and US9. In cells transfected with individual tegument proteins, this matrix is less prevalent. Similarly, coprecipitation of gE/gI and US9 was observed in HSV-infected cells but not in transfected cells, which argued against direct US9-gE/gI interactions. These studies suggest that gE/gI and US9 binding to these tegument proteins has neuron specific effects on virus HSV assembly, a process required for axonal transport of enveloped particles.IMPORTANCE Herpes simplex viruses 1 and 2 and varicella-zoster virus cause significant morbidity and mortality. One basic property of these viruses is the capacity to establish latency in the sensory neurons and to reactivate from latency and then cause disease in peripheral tissues, such as skin and mucosal epithelia. The transport of nascent HSV particles from neuron cell bodies into axons and along axons to axon tips in the periphery is an important component of this reactivation and reinfection. Two HSV membrane proteins, gE/gI and US9, play an essential role in these processes. Our studies help elucidate how HSV gE/gI and US9 promote the assembly of virus particles and sorting of these virions into neuronal axons.