title: A facile quantitative assay for viral particle genesis reveals cooperativity in virion assembly and saturation of an antiviral protein.
abstract: Conventional assays of viral particle assembly and release are time consuming and laborious. We have developed an enzymatic virus-like particle (VLP) genesis assay that rapid and quantitative and is also versatile and applicable to diverse viruses including HIV-1 and Ebola virus. Using this assay, which has a dynamic range of several orders of magnitude, we show that the efficiency of VLP assembly and release, i.e., the fraction of the expressed protein that is assembled into extracellular particles, is dependent on the absolute level of expression of either HIV-1 Gag or Ebola virus VP40. We also demonstrate that the activity of the antiviral factor tetherin is dependent on the level of HIV-1 Gag expression and the numbers of VLPs generated, and appears to become saturated as these parameters are increased.
authors.affiliation: Howard Hughes Medical Institute, Laboratory of Retrovirology, Aaron Diamond AIDS Research Center, The Rockefeller University, 455 First Avenue, New York, NY 10016, USA.
authors.forename: Shalini S
authors.initials: SS
authors.lastname: Yadav
authors.forename: Sam J
authors.initials: SJ
authors.lastname: Wilson
authors.forename: Paul D
authors.initials: PD
authors.lastname: Bieniasz
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D002460:Cell Line; D029043:Ebolavirus / Q000472:pathogenicity*; D058851:GPI-Linked Proteins / Q000378:metabolism; D015497:HIV-1 / Q000472:pathogenicity*; D054884:Host-Pathogen Interactions; D006801:Humans; D009698:Nucleoproteins / Q000378:metabolism; D014758:Viral Core Proteins / Q000378:metabolism; D014771:Virion / Q000378:metabolism*; D019065:Virus Assembly; D057074:Virus Release; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000378:metabolism; D000074010:Bone Marrow Stromal Antigen 2
chemical_list: D015703:Antigens, CD; D058851:GPI-Linked Proteins; D009698:Nucleoproteins; D014758:Viral Core Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; C094572:BST2 protein, human; D000074010:Bone Marrow Stromal Antigen 2; C084819:nucleoprotein VP40, Ebola virus