Visualizing Association of the Retroviral Gag Protein with Unspliced Viral RNA in the Nucleus.
Gag proteins
RNA trafficking
Rous sarcoma virus
genomic RNA packaging
live cell imaging
nucleocytoplasmic trafficking
retrovirus assembly
Journal
mBio
ISSN: 2150-7511
Titre abrégé: mBio
Pays: United States
ID NLM: 101519231
Informations de publication
Date de publication:
07 04 2020
07 04 2020
Historique:
entrez:
9
4
2020
pubmed:
9
4
2020
medline:
18
2
2021
Statut:
epublish
Résumé
Packaging of genomic RNA (gRNA) by retroviruses is essential for infectivity, yet the subcellular site of the initial interaction between the Gag polyprotein and gRNA remains poorly defined. Because retroviral particles are released from the plasma membrane, it was previously thought that Gag proteins initially bound to gRNA in the cytoplasm or at the plasma membrane. However, the Gag protein of the avian retrovirus Rous sarcoma virus (RSV) undergoes active nuclear trafficking, which is required for efficient gRNA encapsidation (L. Z. Scheifele, R. A. Garbitt, J. D. Rhoads, and L. J. Parent, Proc Natl Acad Sci U S A 99:3944-3949, 2002, https://doi.org/10.1073/pnas.062652199; R. Garbitt-Hirst, S. P. Kenney, and L. J. Parent, J Virol 83:6790-6797, 2009, https://doi.org/10.1128/JVI.00101-09). These results raise the intriguing possibility that the primary contact between Gag and gRNA might occur in the nucleus. To examine this possibility, we created a RSV proviral construct that includes 24 tandem repeats of MS2 RNA stem-loops, making it possible to track RSV viral RNA (vRNA) in live cells in which a fluorophore-conjugated MS2 coat protein is coexpressed. Using confocal microscopy, we observed that both wild-type Gag and a nuclear export mutant (Gag.L219A) colocalized with vRNA in the nucleus. In live-cell time-lapse images, the wild-type Gag protein trafficked together with vRNA as a single ribonucleoprotein (RNP) complex in the nucleoplasm near the nuclear periphery, appearing to traverse the nuclear envelope into the cytoplasm. Furthermore, biophysical imaging methods suggest that Gag and the unspliced vRNA physically interact in the nucleus. Taken together, these data suggest that RSV Gag binds unspliced vRNA to export it from the nucleus, possibly for packaging into virions as the viral genome.
Identifiants
pubmed: 32265329
pii: mBio.00524-20
doi: 10.1128/mBio.00524-20
pmc: PMC7157774
pii:
doi:
Substances chimiques
Gene Products, gag
0
RNA, Viral
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : NCI NIH HHS
ID : F31 CA171862
Pays : United States
Organisme : NCI NIH HHS
ID : F31 CA196292
Pays : United States
Organisme : NCI NIH HHS
ID : F30 CA214010
Pays : United States
Organisme : NIAID NIH HHS
ID : P50 AI150464
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA076534
Pays : United States
Organisme : NIH HHS
ID : S10 OD010756
Pays : United States
Organisme : NIGMS NIH HHS
ID : P50 GM103297
Pays : United States
Informations de copyright
Copyright © 2020 Maldonado et al.
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