title: HSV-1 Glycoproteins Are Delivered to Virus Assembly Sites Through Dynamin-Dependent Endocytosis.
abstract: Herpes simplex virus-1 (HSV-1) is a large enveloped DNA virus that belongs to the family of Herpesviridae. It has been recently shown that the cytoplasmic membranes that wrap the newly assembled capsids are endocytic compartments derived from the plasma membrane. Here, we show that dynamin-dependent endocytosis plays a major role in this process. Dominant-negative dynamin and clathrin adaptor AP180 significantly decrease virus production. Moreover, inhibitors targeting dynamin and clathrin lead to a decreased transport of glycoproteins to cytoplasmic capsids, confirming that glycoproteins are delivered to assembly sites via endocytosis. We also show that certain combinations of glycoproteins colocalize with each other and with the components of clathrin-dependent and -independent endocytosis pathways. Importantly, we demonstrate that the uptake of neutralizing antibodies that bind to glycoproteins when they become exposed on the cell surface during virus particle assembly leads to the production of non-infectious HSV-1. Our results demonstrate that transport of viral glycoproteins to the plasma membrane prior to endocytosis is the major route by which these proteins are localized to the cytoplasmic virus assembly compartments. This highlights the importance of endocytosis as a major protein-sorting event during HSV-1 envelopment.
authors.affiliation: Division of Virology, Department of Pathology, Cambridge University, Cambridge, CB2 1QP, UK.
authors.forename: Anna
authors.initials: A
authors.lastname: Albecka
authors.affiliation: Laser Analytics Group, Department of Chemical Engineering and Biotechnology, Cambridge University, Cambridge, CB2 3RA, UK.
authors.forename: Romain F
authors.initials: RF
authors.lastname: Laine
authors.affiliation: Division of Virology, Department of Pathology, Cambridge University, Cambridge, CB2 1QP, UK.
authors.forename: Anne F J
authors.initials: AF
authors.lastname: Janssen
authors.affiliation: Laser Analytics Group, Department of Chemical Engineering and Biotechnology, Cambridge University, Cambridge, CB2 3RA, UK.
authors.forename: Clemens F
authors.initials: CF
authors.lastname: Kaminski
authors.affiliation: Division of Virology, Department of Pathology, Cambridge University, Cambridge, CB2 1QP, UK.
authors.forename: Colin M
authors.identifier: http://orcid.org/0000-0001-9918-9998
authors.initials: CM
authors.lastname: Crump
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D002966:Clathrin / Q000378:metabolism; D034281:Dynamins / Q000378:metabolism*; D004705:Endocytosis; D006023:Glycoproteins / Q000378:metabolism*; D018259:Herpesvirus 1, Human / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D034041:Monomeric Clathrin Assembly Proteins / Q000378:metabolism; D021381:Protein Transport; D014709:Vero Cells; D014764:Viral Proteins / Q000378:metabolism*; D019065:Virus Assembly
chemical_list: D002966:Clathrin; D006023:Glycoproteins; D034041:Monomeric Clathrin Assembly Proteins; D014764:Viral Proteins; C439023:clathrin assembly protein AP180; D034281:Dynamins
keywords: HSV; dSTORM; dynamin; endocytosis; glycoprotein; virus assembly