Isolation and preliminary characterization of herpes simplex virus 1 primary enveloped virions from the perinuclear space.
Herpes simplex virus 1 (HSV-1) nucleocapsids exit the nucleus by budding into the inner nuclear membrane, where they exist briefly as primary enveloped virions. These virus particles subsequently fuse their envelopes with the outer nuclear membrane, permitting nucleocapsids to then enter the cytoplasm and complete assembly. We have developed a method to isolate primary enveloped virions from HSV-1-infected cells and subjected the primary enveloped virion preparation to MALDI-MS/MS (matrix-assisted laser desorption ionization-tandem mass spectrometry) analyses. We identified most capsid proteins, a tegument protein (VP22), a glycoprotein (gD), and a cellular protein (annexin A2) in the primary enveloped virion preparation. We determined that annexin A2 does not play an essential role in infection under our experimental conditions. Elucidating the structure and biochemical properties of this unique virus assembly intermediate will provide new insights into HSV-1 biology.
Padula
Maryn E
ME
Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, USA.
Sydnor
Mariam L
ML
Wilson
Duncan W
DW
D000818:Animals; D017306:Annexin A2 / Q000302:isolation & purification; D019556:COS Cells; D036022:Capsid Proteins / Q000302:isolation & purification*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004279:DNA, Viral / Q000032:analysis; D018259:Herpesvirus 1, Human / Q000302:isolation & purification* / Q000378:metabolism; D009685:Nuclear Envelope / Q000821:virology; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry; D014759:Viral Envelope Proteins / Q000302:isolation & purification*; D014771:Virion / Q000302:isolation & purification* / Q000378:metabolism
D017306:Annexin A2; D036022:Capsid Proteins; D004279:DNA, Viral; D014759:Viral Envelope Proteins