title: Structural analysis of herpes simplex virus by optical super-resolution imaging.
abstract: 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.
authors.affiliation: Laser Analytics Group, Department of Chemical Engineering and Biotechnology, Cambridge University, Pembroke Street, Cambridge CB2 3RA, UK.
authors.forename: Romain F
authors.initials: RF
authors.lastname: Laine
authors.affiliation: Division of Virology, Department of Pathology, Cambridge University, Tennis Court Road, Cambridge CB2 1QP, UK.
authors.forename: Anna
authors.initials: A
authors.lastname: Albecka
authors.affiliation: Department of Biotechnology and Biophysics, Julius-Maximilians-University, Am Hubland, D-97074 Würzburg, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: van de Linde
authors.affiliation: Laser Analytics Group, Department of Chemical Engineering and Biotechnology, Cambridge University, Pembroke Street, Cambridge CB2 3RA, UK.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Rees
authors.affiliation: Division of Virology, Department of Pathology, Cambridge University, Tennis Court Road, Cambridge CB2 1QP, UK.
authors.forename: Colin M
authors.initials: CM
authors.lastname: Crump
authors.affiliation: Laser Analytics Group, Department of Chemical Engineering and Biotechnology, Cambridge University, Pembroke Street, Cambridge CB2 3RA, UK.
authors.forename: Clemens F
authors.initials: CF
authors.lastname: Kaminski
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D002460:Cell Line; D020285:Cryoelectron Microscopy; D005260:Female; D018259:Herpesvirus 1, Human; D006801:Humans; D007091:Image Processing, Computer-Assisted; D015603:Keratinocytes / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D008853:Microscopy / Q000379:methods*; D008856:Microscopy, Fluorescence; D061848:Optical Imaging / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry; D018139:Simplexvirus / Q000648:ultrastructure*; D014759:Viral Envelope Proteins / Q000737:chemistry; D014764:Viral Proteins / Q000737:chemistry; D015678:Viral Structural Proteins / Q000737:chemistry; D014771:Virion / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins