title: Direct evidence for intracellular anterograde co-transport of M-PMV Gag and Env on microtubules.
abstract: The intracellular transport of Mason-Pfizer monkey virus (M-PMV) assembled capsids from the pericentriolar region to the plasma membrane (PM) requires trafficking of envelope glycoprotein (Env) to the assembly site via the recycling endosome. However, it is unclear if Env-containing vesicles play a direct role in trafficking capsids to the PM. Using live cell microscopy, we demonstrate, for the first time, anterograde co-transport of Gag and Env. Nocodazole disruption of microtubules had differential effects on Gag and Env trafficking, with pulse-chase assays showing a delayed release of Env-deficient virions. Particle tracking demonstrated an initial loss of linear movement of GFP-tagged capsids and mCherry-tagged Env, followed by renewed movement of Gag but not Env at 4h post-treatment. Thus, while delayed capsid trafficking can occur in the absence of microtubules, efficient anterograde transport of capsids appears to be mediated by microtubule-associated Env-containing vesicles.
authors.affiliation: Emory Vaccine Center, Yerkes National Primate Research Center, 954 Gatewood Road NE, Atlanta, GA 30329, USA. Electronic address: vvg3@cdc.gov.
authors.forename: Lara E
authors.initials: LE
authors.lastname: Pereira
authors.affiliation: Emory Vaccine Center, Yerkes National Primate Research Center, 954 Gatewood Road NE, Atlanta, GA 30329, USA. Electronic address: jrainey@emory.edu.
authors.forename: Jasmine
authors.initials: J
authors.lastname: Clark
authors.affiliation: Department of Biochemistry and Microbiology, Institute of Chemical Technology, Technicka 3, 166 28 Prague, Czech Republic. Electronic address: Petra.Grznarova@vscht.cz.
authors.forename: Petra
authors.initials: P
authors.lastname: Grznarova
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, 2015 Uppergate Drive, Atlanta, GA 30322, USA. Electronic address: xiaoyun.wen@emory.edu.
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Wen
authors.affiliation: Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, MT, USA. Electronic address: RLaCasse@niaid.nih.gov.
authors.forename: Rachel
authors.initials: R
authors.lastname: LaCasse
authors.affiliation: Department of Biochemistry and Microbiology, Institute of Chemical Technology, Technicka 3, 166 28 Prague, Czech Republic. Electronic address: Tomas.Ruml@vscht.cz.
authors.forename: Tomas
authors.initials: T
authors.lastname: Ruml
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, 2015 Uppergate Drive, Atlanta, GA 30322, USA. Electronic address: paul.spearman@emory.edu.
authors.forename: Paul
authors.initials: P
authors.lastname: Spearman
authors.affiliation: Emory Vaccine Center, Yerkes National Primate Research Center, 954 Gatewood Road NE, Atlanta, GA 30329, USA. Electronic address: eric.hunter2@emory.edu.
authors.forename: Eric
authors.initials: E
authors.lastname: Hunter
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000821:virology; D002522:Chlorocebus aethiops; D015686:Gene Products, env / Q000235:genetics / Q000378:metabolism*; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism*; D008253:Macaca mulatta; D016093:Mason-Pfizer monkey virus / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism / Q000821:virology*; D021381:Protein Transport; D016097:Simian Acquired Immunodeficiency Syndrome / Q000378:metabolism / Q000821:virology*
chemical_list: D015686:Gene Products, env; D015683:Gene Products, gag
keywords: Anterograde transport; Cytoskeleton; Envelope; Gag; Live cell-imaging; M-PMV