title: Glycoprotein M of herpes simplex virus 1 is incorporated into virions during budding at the inner nuclear membrane.
abstract: It is widely accepted that nucleocapsids of herpesviruses bud through the inner nuclear membrane (INM), but few studies have been undertaken to characterize the composition of these nascent virions. Such knowledge would shed light on the budding reaction at the INM and subsequent steps in the egress pathway. The present study focuses on glycoprotein M (gM), a type III integral membrane protein of herpes simplex virus 1 (HSV-1) that likely contains eight transmembrane domains. The results indicated that gM localized primarily at the perinuclear region, with especially bright staining near the nuclear membrane (NM). Immunogold electron microscopic analysis indicated that, like gB and gD (M. R. Torrisi et al., J. Virol. 66:554-561, 1992), gM localized within both leaflets of the NM, the envelopes of nascent virions that accumulate in the perinuclear space, and the envelopes of cytoplasmic and mature extracellular virus particles. Indirect immunofluorescence studies revealed that gM colocalized almost completely with a marker of the Golgi apparatus and partially with a marker of the trans-Golgi network (TGN), whether or not these markers were displaced to the perinuclear region during infection. gM was also located in punctate extensions and invaginations of the NM induced by the absence of a viral kinase encoded by HSV-1 U(S)3 and within virions located in these extensions. Our findings therefore support the proposition that gM, like gB and gD, becomes incorporated into the virion envelope upon budding through the INM. The localization of viral glycoproteins and Golgi and TGN markers to a perinuclear region may represent a mechanism to facilitate the production of infectious nascent virions, thereby increasing the amount of infectivity released upon cellular lysis.
authors.affiliation: C5169 Veterinary Education Center, Cornell University, Ithaca, NY 14853, USA. jdb11@cornell.edu
authors.forename: Joel D
authors.initials: JD
authors.lastname: Baines
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Wills
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Jacob
authors.forename: Janice
authors.initials: J
authors.lastname: Pennington
authors.forename: Bernard
authors.initials: B
authors.lastname: Roizman
mesh_terms: D002460:Cell Line; D019084:Fluorescent Antibody Technique, Indirect; D017353:Gene Deletion; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity* / Q000648:ultrastructure; D006801:Humans; D016253:Microscopy, Immunoelectron; D009685:Nuclear Envelope / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D014759:Viral Envelope Proteins