Physical trapping of HIV-1 synaptic complex by different structural classes of integrase strand transfer inhibitors.
Raltegravir is an FDA approved inhibitor directed against human immunodeficiency virus type 1 (HIV-1) integrase (IN). In this study, we investigated the mechanisms associated with multiple strand transfer inhibitors capable of inhibiting concerted integration by HIV-1 IN. The results show raltegravir, elvitegravir, MK-2048, RDS 1997, and RDS 2197 all appear to encompass a common inhibitory mechanism by modifying IN-viral DNA interactions. These structurally different inhibitors bind to and inactivate the synaptic complex, an intermediate in the concerted integration pathway in vitro. The inhibitors physically trap the synaptic complex, thereby preventing target DNA binding and thus concerted integration. The efficiency of a particular inhibitor to trap the synaptic complex observed on native agarose gels correlated with its potency for inhibiting the concerted integration reaction, defined by IC(50) values for each inhibitor. At low nanomolar concentrations (<50 nM), raltegravir displayed a time-dependent inhibition of concerted integration, a property associated with slow-binding inhibitors. Studies of raltegravir-resistant IN mutants N155H and Q148H without inhibitors demonstrated that their capacity to assemble the synaptic complex and promote concerted integration was similar to their reported virus replication capacities. The concerted integration activity of Q148H showed a higher cross-resistance to raltegravir than observed with N155H, providing evidence as to why the Q148H pathway with secondary mutations is the predominant pathway upon prolonged treatment. Notably, MK-2048 is equally potent against wild-type IN and raltegravir-resistant IN mutant N155H, suggesting this inhibitor may bind similarly within their drug-binding pockets.
Pandey
Krishan K
KK
Institute for Molecular Virology, Saint Louis University Health Sciences Center, 1100 South Grand Boulevard, Saint Louis, Missouri 63104, USA. pandeykk@slu.edu
Bera
Sibes
S
Vora
Ajaykumar C
AC
Grandgenett
Duane P
DP
D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism; D006801:Humans; D019429:Integrase Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D019426:Integrases / Q000235:genetics / Q000494:pharmacology / Q000627:therapeutic use; D009154:Mutation / Q000187:drug effects; D011760:Pyrrolidinones / Q000494:pharmacology; D015363:Quinolones / Q000494:pharmacology; D000068898:Raltegravir Potassium; D014779:Virus Replication / Q000187:drug effects / Q000235:genetics
D004279:DNA, Viral; D019429:Integrase Inhibitors; D011760:Pyrrolidinones; D015363:Quinolones; D000068898:Raltegravir Potassium; C509700:elvitegravir; D019426:Integrases
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