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Cited by 345HIV Research and TreatmentHIV/AIDS drug development and treatmentVirus-based gene therapy research

Retroviral DNA integration directed by HIV integration protein in vitro

Open access
Bushman, Frederic D., Fujiwara, Tamio, Craigie, Robert·1990·Science
DOI 10.1126/science.2171144
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Abstract

Efficient retroviral growth requires integration of a DNA copy of the viral RNA genome into a chromosome of the host. As a first step in analyzing the mechanism of integration of human immunodeficiency virus (HIV) DNA, a cell-free system was established that models the integration reaction. The in vitro system depends on the HIV integration (IN) protein, which was partially purified from insect cells engineered to express IN protein in large quantities. Integration was detected in a biological assay that scores the insertion of a linear DNA containing HIV terminal sequences into a λ DNA target. Some integration products generated in this assay contained five-base pair duplications of the target DNA at the recombination junctions, a characteristic of HIV integration in vivo; the remaining products contained aberrant junctional sequences that may have been produced in a variation of the normal reaction. These results indicate that HIV IN protein is the only viral protein required to insert model HIV DNA sequences into a target DNA in vitro.

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Crossref · 10.1126/science.2171144

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  • The cis-acting att sites are present as imperfect inverted repeats and are both necessary and sufficient for integration in vitro and in vivo.

    “These cis-acting att sites are present in the form of imperfect inverted repeats and are both necessary and sufficient for integration both in vitro and in vivo (Fig. 7) (Colicelli and Goff 1988, Katzman et al. 1989, Bushman et al. 1990, Murphy et al. 1993, Brown et al. 1999).”

    From Beyond reverse transcription: molecular mechanisms and emerging paradigms in retroviral replication (2026) · cites this paper as Bushman et al. 1990

  • PIC-containing reverse-transcribed dsDNA can be integrated into heterologous DNA in vitro.

    “These studies demonstrated that PIC-containing reverse-transcribed dsDNA could be integrated into a heterologous DNA (Fujiwara and Mizuuchi 1988, Fujiwara and Craigie 1989, Bushman et al. 1990, Craigie et al. 1990, Katz et al. 1990, Lee and Coffin 1990, Engelman et al. 1991, Brown 1997).”

    From Beyond reverse transcription: molecular mechanisms and emerging paradigms in retroviral replication (2026) · cites this paper as Bushman et al. 1990

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Authors
Bushman, F. D., Fujiwara, T., & Craigie, R.
Affiliations
National Institutes of Health (US)
ORCID
Venue
Science
Publisher
American Association for the Advancement of Science (AAAS)
Published
1990-09-28
Volume
249
Issue
4976
Pages
1555-1558
ISSN
  • 0036-8075 (ISSN-L)
  • 1095-9203
DOI
10.1126/science.2171144
OpenAlex
W2050569727
Cited by
338
References
26
Citations by year
2026: 1 · 2025: 1 · 2023: 3 · 2022: 3 · 2021: 7 · 2020: 1 · 2019: 5 · 2018: 2 · 2017: 5 · 2016: 7 · 2015: 11 · 2014: 12 · 2013: 8 · 2012: 9
FWCI
6.48
Type
journal article
Language
en
Open access
Licence
public-domain
Locations
  • Science (publishedVersion)
  • PubMed (publishedVersion)
  • Zenodo (CERN European Organization for Nuclear Research) (submittedVersion, public-domain)
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
retroviral integration, HIV-1, terminal repeats, recombination junctions, site-specific integration
MeSH terms
Animals, Base Sequence, Cell Line, Cell-Free System, DNA Transposable Elements, DNA Nucleotidyltransferases, DNA, Viral, HIV, Insect Viruses, Models, Structural, Molecular Sequence Data, Leukemia Virus, Murine, Nucleic Acid Conformation, Restriction Mapping, Oligonucleotide Probes, Integrases

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