Consensus HIV-1 subtype A integrase and its raltegravir-resistant variants: Design and characterization of the enzymatic properties

Shadrina, Olga; Krotova, Olga; Agapkina, Julia; Knyazhanskaya, Ekaterina; Korolev, S. P.; Starodubova, Elizaveta; Viklund, Alecia; Lukashov, Vladimir V.; Magnani, Mauro; Medstrand, Patrik; Карпов, В.Л.; Gottikh, Marina; Isaguliants, Maria

doi:10.1016/j.biochi.2014.02.013

Cited by 6 publications

(10 citation statements)

References 56 publications

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“…We used a 21-mer DNA duplex U5B/U5A mimicking the U5 region of HIV-1 DNA (U5-substrate) and the conditions (enzyme and DNA concentrations, buffer composition) described earlier for the analysis of catalytic activities of IN A and IN B [ 22 ] in the 3′-processing reaction.…”

Section: Resultsmentioning

confidence: 99%

“…In particular, information on resistance mutations caused by IN inhibitors in HIV-1 strain FSU-A is limited. To assess the impact of drug resistance mutations on the enzymatic properties of IN of HIV-1 subtype A, we prepared a consensus IN of the FSU-A strain, where RAL- and EVG-resistance mutations were introduced by site-directed mutagenesis [ 21 , 22 ]. The consensus IN sequence of HIV-1 strain FSU-A (INA) differs from the sequence of the best studied IN of HIV-1 subtype B (HXB-2) by substitutions of 16 amino acid residues, nine of which are located in the catalytic domain.…”

Section: Introductionmentioning

confidence: 99%

“…The consensus IN sequence of HIV-1 strain FSU-A (INA) differs from the sequence of the best studied IN of HIV-1 subtype B (HXB-2) by substitutions of 16 amino acid residues, nine of which are located in the catalytic domain. We characterized the catalytic activity of INA and its variants containing two major combinations of RALand EVG-resistance mutations: E92Q, V151I, N155H, G163R, L74M (mutant 1), and Q148K, E138K, G140S (mutant 2) [ 22 ]. The consensus enzyme was significantly more active than IN of subtype B (INB) in 3’-processing and strand transfer reactions.…”

Section: Introductionmentioning

confidence: 99%

“…The consensus enzyme was significantly more active than IN of subtype B (INB) in 3’-processing and strand transfer reactions. The introduction of these mutations significantly increased INA resistance to RAL and EVG but dramatically reduced its catalytic activity in both reactions [ 22 ].…”

Section: Introductionmentioning

confidence: 99%

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Influence of Drug Resistance Mutations on the Activity of HIV-1 Subtypes A and B Integrases: a Comparative Study

et al. 2015

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Integration of human immunodeficiency virus (HIV-1) DNA into the genome of an infected cell is one of the key steps in the viral replication cycle. The viral enzyme integrase (IN), which catalyzes the integration, is an attractive target for the development of new antiviral drugs. However, the HIV-1 therapy often results in the IN gene mutations inducing viral resistance to integration inhibitors. To assess the impact of drug resistance mutations on the activity of IN of HIV-1 subtype A strain FSU-A, which is dominant in Russia, variants of the consensus IN of this subtype containing the primary resistance mutations G118R and Q148K and secondary compensatory substitutions E138K and G140S were prepared and characterized. Comparative study of these enzymes with the corresponding mutants of IN of HIV-1 subtype B strains HXB-2 was performed. The mutation Q148K almost equally reduced the activity of integrases of both subtypes. Its negative effect was partially compensated by the secondary mutations E138K and G140S. Primary substitution G118R had different influence on the activity of proteins of the subtypes A and B, and the compensatory effect of the secondary substitution E138K also depended on the viral subtype. Comparison of the mutants resistance to the known strand transfer inhibitors raltegravir and elvitegravir, and a new inhibitor XZ-259 (a dihydro-1H-isoindol derivative), showed that integrases of both subtypes with the Q148K mutation were insensitive to raltegravir and elvitegravir but were effectively inhibited by XZ-259. The substitution G118R slightly reduced the efficiency of IN inhibition by raltegravir and elvitegravir and caused no resistance to XZ_259.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Influence of Drug Resistance Mutations on the Activity of HIV-1 Subtypes A and B Integrases: a Comparative Study

et al. 2015

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“…A third activity of IN, requiring the full length protein, has been identified by several independent groups and consists in a specific endonucleolytic activity of IN onto a short ODN mimicking the palindromic sequence found at the LTR-LTR junction of 2-LTRc (Delelis et al, 2005, 2007; Shadrina et al, 2014; Zhang et al, 2014). This reaction occurs symmetrically on the two DNA strands, at the CA position involved in the 3′-P reaction.…”

Section: Integrase and Its Catalytic Activitiesmentioning

confidence: 99%

Different Pathways Leading to Integrase Inhibitors Resistance

2017

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Integrase strand-transfer inhibitors (INSTIs), such as raltegravir (RAL), elvitegravir, or dolutegravir (DTG), are efficient antiretroviral agents used in HIV treatment in order to inhibit retroviral integration. By contrast to RAL treatments leading to well-identified mutation resistance pathways at the integrase level, recent clinical studies report several cases of patients failing DTG treatment without clearly identified resistance mutation in the integrase gene raising questions for the mechanism behind the resistance. These compounds, by impairing the integration of HIV-1 viral DNA into the host DNA, lead to an accumulation of unintegrated circular viral DNA forms. This viral DNA could be at the origin of the INSTI resistance by two different ways. The first one, sustained by a recent report, involves 2-long terminal repeat circles integration and the second one involves expression of accumulated unintegrated viral DNA leading to a basal production of viral particles maintaining the viral information.

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Relationship between HIV integrase polymorphisms and integrase inhibitor susceptibility: An in silico analysis

Hutapea

Maladan

Widodo

2018

Heliyon

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Integrase (IN) plays an essential role in HIV-1 replication, by mediating integration of the viral genome into the host cell genome. IN is a potential target of antiretroviral (ARV) therapeutic drugs such as ALLINI, Raltegravir (RAL), and Elvitegravir (EVG). The effect of IN polymorphisms on its structure and binding affinity to the integrase inhibitors (INIs) is not well understood. The goal of this study was to examine the effect of IN polymorphisms on its tertiary structure and binding affinities to INIs using computational approaches. HIV genomes were isolated from patient blood and the IN gene was sequenced to identify polymorphisms. Protein structures were derived using FoldX and the binding affinity of IN for ALLINI, RAL, and EVG was evaluated using a molecular docking method. The binding affinities of ALLINI and EVG for wild-type IN were lower as compared to an IN variant; in contrast, the binding affinity of RAL for the IN variant was lower as compared to wild-type IN. These results suggested that IN variant interacts with ALLINI and EVG more efficiently as compared to the wildtype, which may not cause resistent to the drugs. In vitro and in vivo studies should be done to validate the findings of this study.

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Consensus HIV-1 subtype A integrase and its raltegravir-resistant variants: Design and characterization of the enzymatic properties

Cited by 6 publications

References 56 publications

Influence of Drug Resistance Mutations on the Activity of HIV-1 Subtypes A and B Integrases: a Comparative Study

Influence of Drug Resistance Mutations on the Activity of HIV-1 Subtypes A and B Integrases: a Comparative Study

Different Pathways Leading to Integrase Inhibitors Resistance

Relationship between HIV integrase polymorphisms and integrase inhibitor susceptibility: An in silico analysis

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