The Novel Nuclear Targeting and BFRF1-Interacting Domains of BFLF2 Are Essential for Efficient Epstein-Barr Virus Virion Release.
Amino Acid Sequence
Cell Line
Cell Nucleus
/ virology
Cytoplasm
/ metabolism
Glutathione Transferase
/ metabolism
HEK293 Cells
HeLa Cells
Herpesvirus 4, Human
/ genetics
Humans
Membrane Proteins
/ chemistry
Models, Molecular
Nuclear Envelope
Nuclear Localization Signals
/ metabolism
Protein Conformation
Sequence Analysis, Protein
Viral Proteins
/ chemistry
Virion
/ metabolism
Virus Release
/ genetics
beta Karyopherins
BFLF2
BFRF1
EBV
Epstein-Barr virus
NLS
herpesviruses
importin beta
nuclear egress
nuclear egress complex
nuclear localization signal
Journal
Journal of virology
ISSN: 1098-5514
Titre abrégé: J Virol
Pays: United States
ID NLM: 0113724
Informations de publication
Date de publication:
17 01 2020
17 01 2020
Historique:
received:
04
09
2019
accepted:
27
10
2019
pubmed:
7
11
2019
medline:
24
7
2020
entrez:
8
11
2019
Statut:
epublish
Résumé
Epstein-Barr virus (EBV) genomic DNA is replicated and packaged into procapsids in the nucleus to form nucleocapsids, which are then transported into the cytoplasm for tegumentation and final maturation. The process is facilitated by the coordination of the viral nuclear egress complex (NEC), which consists of BFLF2 and BFRF1. By expression alone, BFLF2 is distributed mainly in the nucleus. However, it colocalizes with BFRF1 at the nuclear rim and in cytoplasmic nuclear envelope-derived vesicles in coexpressing cells, suggesting temporal control of the interaction between BFLF2 and BFRF1 is critical for their proper function. The N-terminal sequence of BFLF2 is less conserved than that of alpha- and betaherpesvirus homologs. Here, we found that BFLF2 amino acids (aa) 2 to 102 are required for both nuclear targeting and its interaction with BFRF1. Coimmunoprecipitation and confocal analysis indicated that aa 82 to 106 of BFLF2 are important for its interaction with BFRF1. Three crucial amino acids (R47, K50, and R52) and several noncontinuous arginine and histidine residues within aa 59 to 80 function together as a noncanonical nuclear localization signal (NLS), which can be transferred onto yellow fluorescent protein (YFP)-LacZ for nuclear targeting in an importin β-dependent manner. Virion secretion is defective in 293 cells harboring a BFLF2 knockout EBV bacmid upon lytic induction and is restored by
Identifiants
pubmed: 31694953
pii: JVI.01498-19
doi: 10.1128/JVI.01498-19
pmc: PMC7000978
pii:
doi:
Substances chimiques
BFRF1 protein, Human herpesvirus 4
0
Membrane Proteins
0
Nuclear Localization Signals
0
Viral Proteins
0
beta Karyopherins
0
Glutathione Transferase
EC 2.5.1.18
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
Copyright © 2020 American Society for Microbiology.
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