Human immunodeficiency virus integration in a cell-free system.
Integration of the viral genome into the nuclear DNA of a host cell plays a pivotal role in the replication of retroviruses. We have developed an in vitro method for studying the biochemistry of human immunodeficiency virus (HIV) integration by using extracts from HIV-infected cells. Analysis of the reaction products showed that HIV integration in vitro accurately reproduces the in vivo process. Integration occurred without apparent specificity for the target sequence, and the integrated provirus was directly flanked by a 5-base-pair duplication of DNA from the target site. HIV integration did not require a high-energy cofactor, and the enzymatic activities required for integration were recovered with the viral DNA when cell extracts were fractionated by gel exclusion chromatography.
Ellison
V
V
Department of Pediatrics, Stanford University Medical Center, California 94305-5428.
Abrams
H
H
Roe
T
T
Lifson
J
J
Brown
P
P
D001483:Base Sequence; D015704:CD4 Antigens / Q000032:analysis; D002460:Cell Line; D002472:Cell Transformation, Viral; D002474:Cell-Free System; D003001:Cloning, Molecular; D004273:DNA, Neoplasm / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D005814:Genes, Viral; D006678:HIV / Q000235:genetics*; D006801:Humans; D008242:Lysogeny; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011533:Proviruses / Q000235:genetics; D013601:T-Lymphocytes
D015704:CD4 Antigens; D004273:DNA, Neoplasm; D004279:DNA, Viral; D015345:Oligonucleotide Probes