A preferred target DNA structure for retroviral integrase in vitro.
The retroviral integrase protein catalyzes the insertion of linear viral DNA ends into the host cell DNA. Although integration in vivo is not site-specific, the detection of local and regional preferences within cellular DNA suggests that the integration reaction can be influenced by specific features of host DNA or chromatin. Here we describe highly preferred in vitro integration sites for avian sarcoma virus and human immunodeficiency virus-1 integrases within the stems of plasmid DNA cruciform structures. The preferred sites are adjacent to the loops in the cruciform and are strand-specific. We suggest that the observed preference is due to the end-like character of the stem loop structure that allows DNA unpairing. From these results we propose that such unpairing may enhance both the processing and the joining steps in the integration reaction, and perhaps other cellular recombination reactions as well.
Katz
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RA
Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111, USA. r_katz@fccc.edu
Gravuer
K
K
Skalka
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AM
D001358:Avian Sarcoma Viruses / Q000201:enzymology*; D001483:Base Sequence; D017931:DNA Primers; D004278:DNA, Superhelical / Q000737:chemistry / Q000378:metabolism; D004279:DNA, Viral / Q000737:chemistry / Q000378:metabolism*; D019427:HIV Integrase / Q000378:metabolism*; D006801:Humans; D019426:Integrases / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D016133:Polymerase Chain Reaction; D013379:Substrate Specificity
D017931:DNA Primers; D004278:DNA, Superhelical; D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides; D019427:HIV Integrase; D019426:Integrases