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
| Autor: | Laura Marler, Xue Zhi Zhao, Stephen H. Hughes, Terrence R. Burke, Christophe Marchand, Evgeny Kiselev, Yves Pommier, Barry C. Johnson, Mathieu Métifiot |
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| Rok vydání: | 2016 |
| Předmět: |
Models
Molecular 0301 basic medicine Guanine Integrase inhibitor HIV Integrase Substrate Specificity 03 medical and health sciences chemistry.chemical_compound Catalytic Domain Drug Resistance Viral Genetics medicine Magnesium HIV Integrase Inhibitors Ions chemistry.chemical_classification 030102 biochemistry & molecular biology biology Nucleic Acid Enzymes Elvitegravir Oligonucleotide Raltegravir Virology 3. Good health Integrase 030104 developmental biology Enzyme chemistry DNA Viral Mutation Dolutegravir Biocatalysis biology.protein DNA medicine.drug |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkw592 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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