Effect of DNA modifications on DNA processing by HIV-1 integrase and inhibitor binding: role of DNA backbone flexibility and an open catalytic site.
Integration of the viral cDNA into host chromosomes is required for viral replication. Human immunodeficiency virus integrase catalyzes two sequential reactions, 3'-processing (3'-P) and strand transfer (ST). The first integrase inhibitors are undergoing clinical trial, but interactions of inhibitors with integrase and DNA are not well understood in the absence of a co-crystal structure. To increase our understanding of integrase interactions with DNA, we examined integrase catalysis with oligonucleotides containing DNA backbone, base, and groove modifications placed at unique positions surrounding the 3'-processing site. 3'-Processing was blocked with substrates containing constrained sugars and alpha-anomeric residues, suggesting that integrase requires flexibility of the phosphodiester backbone at the 3'-P site. Of several benzo[a]pyrene 7,8-diol 9,10-epoxide (BaP DE) adducts tested, only the adduct in the minor groove at the 3'-P site inhibited 3'-P, suggesting the importance of the minor groove contacts for 3'-P. ST occurred in the presence of bulky BaP DE DNA adducts attached to the end of the viral DNA suggesting opening of the active site for ST. Position-specific effects of these BaP DE DNA adducts were found for inhibition of integrase by diketo acids. Together, these results demonstrate the importance of DNA structure and specific contacts with the viral DNA processing site for inhibition by integrase inhibitors.
Johnson
Allison A
AA
Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institutes of Health/DHHS, Bethesda, MD 20892, USA.
Sayer
Jane M
JM
Yagi
Haruhiko
H
Patil
Sachindra S
SS
Debart
Françoise
F
Maier
Martin A
MA
Corey
David R
DR
Vasseur
Jean-Jacques
JJ
Burke
Terrence R
TR
Marquez
Victor E
VE
Jerina
Donald M
DM
Pommier
Yves
Y
D001483:Base Sequence; D001665:Binding Sites; D002384:Catalysis; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D018736:DNA Adducts / Q000737:chemistry / Q000378:metabolism; D004279:DNA, Viral / Q000737:chemistry / Q000378:metabolism; D004101:Dihydroxydihydrobenzopyrenes / Q000737:chemistry / Q000378:metabolism; D004852:Epoxy Compounds / Q000737:chemistry / Q000378:metabolism; D019427:HIV Integrase / Q000378:metabolism*; D019428:HIV Integrase Inhibitors / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D015497:HIV-1 / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D008956:Models, Chemical; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism; D018583:Pliability; D013379:Substrate Specificity; D016662:Virus Integration
D018736:DNA Adducts; D004279:DNA, Viral; D004101:Dihydroxydihydrobenzopyrenes; D004852:Epoxy Compounds; D019428:HIV Integrase Inhibitors; D009841:Oligonucleotides; D004247:DNA; D019427:HIV Integrase