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

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Christensen, Devin E., Ganser-Pornillos, Barbie K., Johnson, Jarrod S., Pornillos, Owen, Sundquist, Wesley I.·2020·Science
DOI 10.1126/science.abc8420PMC PMC8022914
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Abstract

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.

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Authors
Christensen, D. E., Ganser-Pornillos, B. K., Johnson, J. S., Pornillos, O., & Sundquist, W. I.
Affiliations
  • University of Utah (US)
  • Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, VA 22903, USA.
  • University of Virginia (US)
Corresponding author
  • Christensen, Devin E.
  • Pornillos, Owen
  • Sundquist, Wesley I.
ORCID
Venue
Science
Publisher
American Association for the Advancement of Science (AAAS)
Published
2020-10-09
Volume
370
Issue
6513
ISSN
  • 0036-8075 (ISSN-L)
  • 1095-9203
DOI
10.1126/science.abc8420
PMC
PMC8022914
OpenAlex
W3092270865
Cited by
150
References
6
Citations by year
2026: 19 · 2025: 24 · 2024: 20 · 2023: 25 · 2022: 25 · 2021: 36 · 2020: 1
FWCI
11.73
Type
journal article
Language
en
Open access
Locations
  • Science (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
National Institute of Allergy and Infectious Diseases (AI150464)
Topics
  • Life Sciences › Immunology and Microbiology › Virology › HIV Research and Treatment
  • Health Sciences › Medicine › Infectious Diseases › HIV/AIDS drug development and treatment
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
Keywords
HIV-1, capsid, reverse transcription, viral integration, preintegration complex, cell-free reconstitution, viral DNA, in situ imaging, viral life cycle
MeSH terms
Capsid, Cell-Free System, Humans, Virus Replication, HIV-1, Virus Integration

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