Laine Romain F, Albecka Anna, van de Linde Sebastian, Rees Eric J, Crump Colin M, Kaminski Clemens F
Laser Analytics Group, Department of Chemical Engineering and Biotechnology, Cambridge University, Pembroke Street, Cambridge CB2 3RA, UK.
Division of Virology, Department of Pathology, Cambridge University, Tennis Court Road, Cambridge CB2 1QP, UK.
Nat Commun. 2015 Jan 22;6:5980. doi: 10.1038/ncomms6980.
Herpes simplex virus type-1 (HSV-1) is one of the most widespread pathogens among humans. Although the structure of HSV-1 has been extensively investigated, the precise organization of tegument and envelope proteins remains elusive. Here we use super-resolution imaging by direct stochastic optical reconstruction microscopy (dSTORM) in combination with a model-based analysis of single-molecule localization data, to determine the position of protein layers within virus particles. We resolve different protein layers within individual HSV-1 particles using multi-colour dSTORM imaging and discriminate envelope-anchored glycoproteins from tegument proteins, both in purified virions and in virions present in infected cells. Precise characterization of HSV-1 structure was achieved by particle averaging of purified viruses and model-based analysis of the radial distribution of the tegument proteins VP16, VP1/2 and pUL37, and envelope protein gD. From this data, we propose a model of the protein organization inside the tegument.
单纯疱疹病毒1型（HSV-1）是人类中传播最广泛的病原体之一。尽管对HSV-1的结构进行了广泛研究，但被膜和包膜蛋白的精确组织仍不清楚。在这里，我们使用直接随机光学重建显微镜（dSTORM）进行超分辨率成像，并结合基于模型的单分子定位数据分析，以确定病毒颗粒内蛋白质层的位置。我们使用多色dSTORM成像解析单个HSV-1颗粒内的不同蛋白质层，并区分纯化病毒粒子和感染细胞中存在的病毒粒子中的包膜锚定糖蛋白与被膜蛋白。通过对纯化病毒进行颗粒平均以及对被膜蛋白VP16、VP1/2和pUL37以及包膜蛋白gD的径向分布进行基于模型的分析，实现了对HSV-1结构的精确表征。根据这些数据，我们提出了被膜内蛋白质组织的模型。
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