Effect of topoisomerase inhibitors on the in vitro HIV DNA integration reaction.
Retroviral growth requires as an obligatory step the integration of a DNA copy of the viral RNA into the genomic DNA of the host. Recombinant human immunodeficiency virus type I (HIV-1) integrase (IN) expressed in Escherichia coli efficiently catalyzes the overall in vitro integration reaction, namely, the processing of the LTR ends and the strand transfer reaction. Using the 3' end of synthetic oligonucleotides which match the termini of the HIV-I U5 LTR as substrate and supercoiled pSP65 DNA as target, we have measured the effect of various topoisomerase inhibitors on the functional activity of the IN protein. Among the various drugs tested, the antitumor drug 2N-Methyl, 9-hydroxyellipticinium (NMHE) displays a marked inhibitory effect on the IN-catalyzed U5 insertion. This effect is related to the DNA binding properties of the drug rather than to a selective effect on the IN protein or the DNA-IN protein complex.
Laboratoire de Pharmacologie des Macomolécules Biologiques, CNRS URA 0147, INSERM U 140, Institut Gustave Roussy, Villejuif, France.
S
S
Carteau
J F
JF
Mouscadet
H
H
Goulaouic
F
F
Subra
C
C
Auclair
D000677:Amsacrine / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D002166:Camptothecin / Q000494:pharmacology; D002227:Carbazoles / Q000494:pharmacology; D004254:DNA Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D004611:Ellipticines / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics; D005047:Etoposide / Q000494:pharmacology; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000201:enzymology* / Q000235:genetics; D019426:Integrases; D007700:Kinetics; D008969:Molecular Sequence Data; D009429:Netropsin / Q000494:pharmacology; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis; D010957:Plasmids; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D013379:Substrate Specificity; D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors
D000970:Antineoplastic Agents; D000998:Antiviral Agents; D002227:Carbazoles; D004611:Ellipticines; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors; D000677:Amsacrine; C061416:oxazolopyridocarbazole; C027767:elliptinium; D009429:Netropsin; D005047:Etoposide; D004254:DNA Nucleotidyltransferases; D019426:Integrases; D002166:Camptothecin