Simultaneous tracking of capsid, tegument, and envelope protein localization in living cells infected with triply fluorescent herpes simplex virus 1.
We report here the construction of a triply fluorescent-tagged herpes simplex virus 1 (HSV-1) expressing capsid protein VP26, tegument protein VP22, and envelope protein gB as fusion proteins with monomeric yellow, red, and cyan fluorescent proteins, respectively. The recombinant virus enabled us to monitor the dynamics of these capsid, tegument, and envelope proteins simultaneously in the same live HSV-1-infected cells and to visualize single extracellular virions with three different fluorescent emissions. In Vero cells infected by the triply fluorescent virus, multiple cytoplasmic compartments were found to be induced close to the basal surfaces of the infected cells (the adhesion surfaces of the infected cells on the solid growth substrate). Major capsid, tegument, and envelope proteins accumulated and colocalized in the compartments, as did marker proteins for the trans-Golgi network (TGN) which has been implicated to be the site of HSV-1 secondary envelopment. Moreover, formation of the compartments was correlated with the dynamic redistribution of the TGN proteins induced by HSV-1 infection. These results suggest that HSV-1 infection causes redistribution of TGN membranes to form multiple cytoplasmic compartments, possibly for optimal secondary envelopment. This is the first real evidence for the assembly of all three types of herpesvirus proteins-capsid, tegument, and envelope membrane proteins-in TGN.
Sugimoto
Ken
K
Division of Viral Infection, Department of Infectious Disease Control, International Research Center for Infectious Diseases, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo, Japan.
Uema
Masashi
M
Sagara
Hiroshi
H
Tanaka
Michiko
M
Sata
Tetsutaro
T
Hashimoto
Yasuhiro
Y
Kawaguchi
Yasushi
Y
D000818:Animals; D015415:Biomarkers; D002213:Capsid / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000378:metabolism; D015686:Gene Products, env / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D021381:Protein Transport; D011817:Rabbits; D021601:trans-Golgi Network / Q000378:metabolism
D015415:Biomarkers; D015686:Gene Products, env