Molecular Impact of the M184V Mutation in Human Immunodeficiency Virus Type 1 Reverse Transcriptase

Diallo, Karidia; Götte, Matthias; Wainberg, Mark A.

doi:10.1128/aac.47.11.3377-3383.2003

Cited by 65 publications

(65 citation statements)

References 72 publications

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“…4A), which results from a G-to-A substitution in an APOBEC3G-favored dinucleotide context. On the other hand, M184V or M184T, which are caused by A-to-G or T-to-C substitutions (27), were never observed. Proviruses with M184I were detected in every single infection with Vif mutants (K22E, 24/34, 70%; E45G, 5/33, 15%; 144AAA, 15/32, 47%) but were notably absent in all WT infections (Fig.…”

Section: Hiv-1 Vif Mutants Lead To the Accumulation Of 3tc-resistant mentioning

confidence: 99%

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Cytidine deamination induced HIV-1 drug resistance

Mulder

Harari²,

Simon³

2008

Proc. Natl. Acad. Sci. U.S.A.

148

169

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The HIV-1 Vif protein is essential for overcoming the antiviral activity of DNA-editing apolipoprotein B mRNA editing enzyme, catalytic polypeptide 3 (APOBEC3) cytidine deaminases. We show that naturally occurring HIV-1 Vif point mutants with suboptimal anti-APOBEC3G activity induce the appearance of proviruses with lamivudine (3TC) drug resistance-associated mutations before any drug exposure. These mutations, ensuing from cytidine deamination events, were detected in >40% of proviruses with partially defective Vif mutants. Transfer of drug resistance from hypermutated proviruses via recombination allowed for 3TC escape under culture conditions prohibitive for any WT viral growth. These results demonstrate that defective hypermutated genomes can shape the phenotype of the circulating viral population. Partially active Vif alleles resulting in incomplete neutralization of cytoplasmic APOBEC3 molecules are directly responsible for the generation of a highly diverse, yet G-to-A biased, proviral reservoir, which can be exploited by HIV-1 to generate viable and drug-resistant progenies.hypermutation ͉ reservoir ͉ diversity ͉ reverse transcription

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Section: Hiv-1 Vif Mutants Lead To the Accumulation Of 3tc-resistant mentioning

confidence: 99%

“…High-level resistance to the nucleoside RT inhibitor 3TC is associated with mutations M184I, M184V, or M184T in the active site of RT (27). A third of all clones sequenced encoded HIV-1 drug-resistance mutation M184I (33%, 44/132 clones; Fig.…”

Section: Hiv-1 Vif Mutants Lead To the Accumulation Of 3tc-resistant mentioning

confidence: 99%

Cytidine deamination induced HIV-1 drug resistance

Mulder

Harari²,

Simon³

2008

Proc. Natl. Acad. Sci. U.S.A.

148

169

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“…RT enzymes were expressed in Escherichia coli BL21 cells (Invitrogen, Burlington, Ontario, Canada) and purified by nickel affinity chromatography and Q Sepharose ion-exchange chromatography, as described previously (33). The polymerase activity of each recombinant RT preparation was evaluated in triplicate by using the synthetic homopolymeric poly(rA)/p(dT) [12][13][14][15][16][17][18] template/primer (T/P) (Midland Certified Reagent Company) as described previously (41). An active unit of RT was defined as the amount of enzyme that incorporates 1 pmol of dTTP in 10 min at 37°C.…”

Section: Chemicalsmentioning

confidence: 99%

“…Kinetics studies were carried out by a modification of a previously described method using homopolymeric poly(rA)/p(dT) [12][13][14][15][16][17][18] and complementary dTTP as the nucleotide substrate (54). The reaction mixture (10 l) contained 50 mM Tris-HCl (pH 7.8), 60 mM KCl, 6 mM MgCl 2 , 5 mM dithiothreitol (DTT), 0.5 U/ml poly(rA)/p(dT) [12][13][14][15][16][17][18] , RT enzymes, variable concentrations of the tracer [ 3 H]dTTP, and cold dTTP (0.2 to 200 M). Reactions were run at 37°C and quenched by the addition of 0.2 ml of 10% cold trichloroacetic acid and 20 mM sodium pyrophosphate to the mixtures; products were filtered onto Millipore 96-well MutiScreen HTS FC filter plates (catalog number MSFCN6B) and sequentially washed with 200 l of 10% TCA and 150 l of 95% ethanol.…”

Section: Chemicalsmentioning

confidence: 99%

“…While RTs containing either the M184V or M184I mutation, associated with resistance to 3TC and FTC, are impaired in the usage of dNTPs (6,15,18), the simultaneous presence of the E138K mutation together with either the M184V or M184I mutation can compensate for this deficit in dNTP usage at low dNTP concentrations through the promotion of tighter dNTP binding. These results indicate that the E138K mutation may also play an important role in the fine-tuning of RT activity.…”

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confidence: 99%

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Subunit-Selective Mutational Analysis and Tissue Culture Evaluations of the Interactions of the E138K and M184I Mutations in HIV-1 Reverse Transcriptase

Oliveira

Quashie

et al. 2012

J Virol

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The emergence of HIV-1 drug resistance remains a major obstacle in antiviral therapy. M184I/V and E138K are signature mutations of clinical relevance in HIV-1 reverse transcriptase (RT) for the nucleoside reverse transcriptase inhibitors (NRTIs) lamivudine (3TC) and emtricitabine (FTC) and the second-generation (new) nonnucleoside reverse transcriptase inhibitor (NNRTI) rilpivirine (RPV), respectively, and the E138K mutation has also been shown to be selected by etravirine in cell culture. The E138K mutation was recently shown to compensate for the low enzyme processivity and viral fitness associated with the M184I/V mutations through enhanced deoxynucleoside triphosphate (dNTP) usage, while the M184I/V mutations compensated for defects in polymerization rates associated with the E138K mutations under conditions of high dNTP concentrations. The M184I mutation was also shown to enhance resistance to RPV and ETR when present together with the E138K mutation. These mutual compensatory effects might also enhance transmission rates of viruses containing these two mutations. Therefore, we performed tissue culture studies to investigate the evolutionary dynamics of these viruses. Through experiments in which E138K-containing viruses were selected with 3TC-FTC and in which M184I/V viruses were selected with ETR, we demonstrated that ETR was able to select for the E138K mutation in viruses containing the M184I/V mutations and that the M184I/V mutations consistently emerged when E138K viruses were selected with 3TC-FTC. We also performed biochemical subunit-selective mutational analyses to investigate the impact of the E138K mutation on RT function and interactions with the M184I mutation. We now show that the E138K mutation decreased rates of polymerization, impaired RNase H activity, and conferred ETR resistance through the p51 subunit of RT, while an enhancement of dNTP usage as a result of the simultaneous presence of both mutations E138K and M184I occurred via both subunits.

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Molecular Mechanisms of Resistance to Nucleoside Reverse Transcriptase Inhibitors

Huigen

Boucher

2009

Antimicrobial Drug Resistance

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Molecular Impact of the M184V Mutation in Human Immunodeficiency Virus Type 1 Reverse Transcriptase

Cited by 65 publications

References 72 publications

Cytidine deamination induced HIV-1 drug resistance

Cytidine deamination induced HIV-1 drug resistance

Subunit-Selective Mutational Analysis and Tissue Culture Evaluations of the Interactions of the E138K and M184I Mutations in HIV-1 Reverse Transcriptase

Molecular Mechanisms of Resistance to Nucleoside Reverse Transcriptase Inhibitors

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