title: Selectivity for strand-transfer over 3'-processing and susceptibility to clinical resistance of HIV-1 integrase inhibitors are driven by key enzyme-DNA interactions in the active site.
abstract: Integrase strand transfer inhibitors (INSTIs) are highly effective against HIV infections. Co-crystal structures of the prototype foamy virus intasome have shown that all three FDA-approved drugs, raltegravir (RAL), elvitegravir and dolutegravir (DTG), act as interfacial inhibitors during the strand transfer (ST) integration step. However, these structures give only a partial sense for the limited inhibition of the 3'-processing reaction by INSTIs and how INSTIs can be modified to overcome drug resistance, notably against the G140S-Q148H double mutation. Based on biochemical experiments with modified oligonucleotides, we demonstrate that both the viral DNA +1 and -1 bases, which flank the 3'-processing site, play a critical role for 3'-processing efficiency and inhibition by RAL and DTG. In addition, the G140S-Q148H (SH) mutant integrase, which has a reduced 3'-processing activity, becomes more active and more resistant to inhibition of 3'-processing by RAL and DTG in the absence of the -1 and +1 bases. Molecular modeling of HIV-1 integrase, together with biochemical data, indicate that the conserved residue Q146 in the flexible loop of HIV-1 integrase is critical for productive viral DNA binding through specific contacts with the virus DNA ends in the 3'-processing and ST reactions. The potency of integrase inhibitors against 3'-processing and their ability to overcome resistance is discussed.
authors.affiliation: Developmental Therapeutics Branch and Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 37 Convent Drive, Bethesda, MD 20892, USA mathieu.metifiot@u-bordeaux.fr.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Métifiot
authors.affiliation: HIV Dynamics and Replication Program, National Cancer Institute at Frederick, Center for Cancer Research, National Institutes of Health, Frederick, MD 21702, USA.
authors.forename: Barry C
authors.initials: BC
authors.lastname: Johnson
authors.affiliation: Developmental Therapeutics Branch and Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 37 Convent Drive, Bethesda, MD 20892, USA.
authors.forename: Evgeny
authors.initials: E
authors.lastname: Kiselev
authors.affiliation: Developmental Therapeutics Branch and Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 37 Convent Drive, Bethesda, MD 20892, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Marler
authors.affiliation: Chemical Biology Laboratory, National Cancer Institute at Frederick, Center for Cancer Research, National Institutes of Health, Frederick, MD 21702, USA.
authors.forename: Xue Zhi
authors.initials: XZ
authors.lastname: Zhao
authors.affiliation: Chemical Biology Laboratory, National Cancer Institute at Frederick, Center for Cancer Research, National Institutes of Health, Frederick, MD 21702, USA.
authors.forename: Terrence R
authors.initials: TR
authors.lastname: Burke
authors.affiliation: Developmental Therapeutics Branch and Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 37 Convent Drive, Bethesda, MD 20892, USA.
authors.forename: Christophe
authors.initials: C
authors.lastname: Marchand
authors.affiliation: HIV Dynamics and Replication Program, National Cancer Institute at Frederick, Center for Cancer Research, National Institutes of Health, Frederick, MD 21702, USA.
authors.forename: Stephen H
authors.initials: SH
authors.lastname: Hughes
authors.affiliation: Developmental Therapeutics Branch and Laboratory of Molecular Pharmacology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 37 Convent Drive, Bethesda, MD 20892, USA pommier@nih.gov.
authors.forename: Yves
authors.initials: Y
authors.lastname: Pommier
mesh_terms: D055162:Biocatalysis / Q000187:drug effects; D020134:Catalytic Domain; D004279:DNA, Viral / Q000378:metabolism*; D024882:Drug Resistance, Viral / Q000187:drug effects*; D006147:Guanine / Q000378:metabolism; D019427:HIV Integrase / Q000737:chemistry / Q000378:metabolism*; D019428:HIV Integrase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D007477:Ions; D008274:Magnesium / Q000494:pharmacology; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D013379:Substrate Specificity / Q000187:drug effects
chemical_list: D004279:DNA, Viral; D019428:HIV Integrase Inhibitors; D007477:Ions; D006147:Guanine; D019427:HIV Integrase; D008274:Magnesium; C514801:p31 integrase protein, Human immunodeficiency virus 1