Reconstitution and visualization of HIV-1 capsid-dependent replication and integration in vitro

During the first half of the viral life cycle, HIV-1 reverse transcribes its RNA genome and integrates the double-stranded DNA copy into a host cell chromosome. Despite progress in characterizing and inhibiting these processes, in situ mechanistic and structural studies remain challenging. This is b...

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Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 2020-10, Vol.370 (6513)
Main Authors: Christensen, Devin E, Ganser-Pornillos, Barbie K, Johnson, Jarrod S, Pornillos, Owen, Sundquist, Wesley I
Format: Article
Language:English
Subjects:
Accumulation
Antiretroviral agents
Archives & records
Capsid - physiology
Capsids
Cell-Free System
Chromosomes
Cloning
Core particles
Cytoplasm
Deoxyribonucleic acid
DNA
Double-stranded RNA
Enzymatic activity
Extrusion
Fullerenes
Gene mapping
Genomes
HIV
HIV-1 - physiology
Human immunodeficiency virus
Humans
Integration
Intermediates
Lattices
Life cycles
Mapping
Mutation
Nuclei (cytology)
Nucleic acids
Nucleotide sequence
Pore formation
Protein arrays
Replication
Reproduction (copying)
Reverse transcription
Ribonucleic acid
RNA
Temporal variations
Uncoating
Virions
Virus Integration
Virus Replication
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Summary:During the first half of the viral life cycle, HIV-1 reverse transcribes its RNA genome and integrates the double-stranded DNA copy into a host cell chromosome. Despite progress in characterizing and inhibiting these processes, in situ mechanistic and structural studies remain challenging. This is because these operations are executed by individual viral preintegration complexes deep within cells. We therefore reconstituted and imaged the early stages of HIV-1 replication in a cell-free system. HIV-1 cores released from permeabilized virions supported efficient, capsid-dependent endogenous reverse transcription to produce double-stranded DNA genomes, which sometimes looped out from ruptured capsid walls. Concerted integration of both viral DNA ends into a target plasmid then proceeded in a cell extract-dependent reaction. This reconstituted system uncovers the role of the capsid in templating replication.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.abc8420