HIV-1 uncoats in the nucleus near sites of integration.
Active Transport, Cell Nucleus
Capsid Proteins
/ analysis
Cell Nucleus
/ virology
Green Fluorescent Proteins
/ analysis
HIV Infections
/ virology
HIV-1
/ physiology
Humans
Nuclear Pore
/ metabolism
Proteolysis
Virus Integration
Virus Replication
Virus Uncoating
mRNA Cleavage and Polyadenylation Factors
/ metabolism
HIV-1
capsid
integration
transcription
uncoating
Journal
Proceedings of the National Academy of Sciences of the United States of America
ISSN: 1091-6490
Titre abrégé: Proc Natl Acad Sci U S A
Pays: United States
ID NLM: 7505876
Informations de publication
Date de publication:
10 03 2020
10 03 2020
Historique:
pubmed:
26
2
2020
medline:
14
7
2020
entrez:
26
2
2020
Statut:
ppublish
Résumé
HIV-1 capsid core disassembly (uncoating) must occur before integration of viral genomic DNA into the host chromosomes, yet remarkably, the timing and cellular location of uncoating is unknown. Previous studies have proposed that intact viral cores are too large to fit through nuclear pores and uncoating occurs in the cytoplasm in coordination with reverse transcription or at the nuclear envelope during nuclear import. The capsid protein (CA) content of the infectious viral cores is not well defined because methods for directly labeling and quantifying the CA in viral cores have been unavailable. In addition, it has been difficult to identify the infectious virions because only one of ∼50 virions in infected cells leads to productive infection. Here, we developed methods to analyze HIV-1 uncoating by direct labeling of CA with GFP and to identify infectious virions by tracking viral cores in living infected cells through viral DNA integration and proviral DNA transcription. Astonishingly, our results show that intact (or nearly intact) viral cores enter the nucleus through a mechanism involving interactions with host protein cleavage and polyadenylation specificity factor 6 (CPSF6), complete reverse transcription in the nucleus before uncoating, and uncoat <1.5 h before integration near (<1.5 μm) their genomic integration sites. These results fundamentally change our current understanding of HIV-1 postentry replication events including mechanisms of nuclear import, uncoating, reverse transcription, integration, and evasion of innate immunity.
Identifiants
pubmed: 32094182
pii: 1920631117
doi: 10.1073/pnas.1920631117
pmc: PMC7071919
doi:
Substances chimiques
Capsid Proteins
0
cleavage factor Im, human
0
mRNA Cleavage and Polyadenylation Factors
0
Green Fluorescent Proteins
147336-22-9
Types de publication
Journal Article
Research Support, N.I.H., Intramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
5486-5493Informations de copyright
Copyright © 2020 the Author(s). Published by PNAS.
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