Compensation by the E138K Mutation in HIV-1 Reverse Transcriptase for Deficits in Viral Replication Capacity and Enzyme Processivity Associated with the M184I/V Mutations

Xu, Hongtao; Asahchop, Eugene L.; Oliveira, Maureen; Quashie, Peter K.; Quan, Yudong; Brenner, Bluma G.; Wainberg, Mark A.

doi:10.1128/jvi.05584-11

Cited by 83 publications

(183 citation statements)

References 40 publications

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“…We found that the E138K, M184I, and M184V mutations each resulted in reduced virion-associated RT activity, and that the E138K/M184I combination restored RT activity to WT levels. Our findings are consistent with results of a more detailed biochemical assessment of HIV-1 RT using purified enzyme preparations that noted the improved processivity of the E138K-M184I/V mutant enzyme compared to that of the WT or M184I/V RT (28). These results provide another example of cross-talk between mutations affecting the nucleoside and nonnucleoside RT inhibitor binding pockets (10,21).…”

Section: Discussionsupporting

confidence: 80%

Interaction of Reverse Transcriptase (RT) Mutations Conferring Resistance to Lamivudine and Etravirine: Effects on Fitness and RT Activity of Human Immunodeficiency Virus Type 1

Hu¹,

Kuritzkes²

2011

J Virol

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Resistance to the nonnucleoside reverse transcriptase inhibitors etravirine and rilpivirine (RPV) is conferred by the E138K mutation in human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT).Clinical trials of RPV administered with lamivudine or emtricitabine showed the emergence of E138K together with M184I, which confers lamivudine and emtricitabine resistance in most patients with virologic failure. To understand why M184I was favored over M184V, we determined the drug susceptibility, infectivity, relative fitness, and reverse transcriptase activity of HIV-1 carrying E138K/M184I or E138K/M184V mutations. Whereas the replication capacity (RC) of the single mutants was reduced compared to that of the wild type (WT), the RC of the two double mutants was comparable to that of the WT in the absence of drug. The RC of the E138K/M184I mutant in the presence of etravirine was significantly greater than that of the E138K and E138K/M184V mutants; the RC of the double mutants was greater than that of the M184I or M184V mutant. Fitness profiles and growth competition experiments showed that the E138K/M184I mutant had a significant replicative advantage over the E138K/M184V mutant in the presence of etravirine and lamivudine. The virion-associated RT activity of the E138K, M184I, or M184V virus was significantly reduced compared to that of the WT, whereas the RT activity of the E138K/M184I virus was significantly greater than that of the WT or E138K/M184V virus. These results suggest that the E138K and M184I/V mutations are mutually compensatory and may explain the frequent occurrence of E138K/M184I after the virologic failure of rilpivirine-, lamivudine-, and emtricitabine-containing regimens.Nonnucleoside reverse transcriptase inhibitors (NNRTIs) are key components of combination antiretroviral therapy. Etravirine (ETV; formerly TMC125) and rilpivirine (RPV; formerly TMC278) are potent next-generation NNRTIs that retain activity against efavirenz (EFV)-resistant viruses carrying the K103N mutation in human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) (1, 3); both are diarylpyrimidine (DAPY) compounds. The E138K mutation, which emerges both in vitro and in vivo, confers resistance to ETV and RPV (2,3,22,23). Although in vitro passage experiments suggested that resistance to ETV and RPV emerges more slowly than resistance to first-generation NNRTIs (e.g., nevirapine [NVP] or EFV) (3, 25), this finding has not been confirmed in clinical trials, which show the emergence of ETV and RPV resistance at the time of virologic failure in most patients treated with these drugs (17, 18).Most antiretroviral regimens include the cytosine analogs lamivudine (3TC) or emtricitabine (FTC) administered as fixed-dose combinations together with zidovudine (ZDV) or abacavir (ABC) in the case of ZDV/3TC and ABC/3TC or with tenofovir (TDF) in the case of TDF/FTC. Resistance to 3TC and FTC is conferred by a valine or isoleucine substitution for the methionine normally found at position 184, which lies in the ...

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Section: Discussionsupporting

confidence: 80%

Interaction of Reverse Transcriptase (RT) Mutations Conferring Resistance to Lamivudine and Etravirine: Effects on Fitness and RT Activity of Human Immunodeficiency Virus Type 1

Hu¹,

Kuritzkes²

2011

J Virol

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“…RNase H activity was assayed by using a 41-mer 5=-32 P-labeled heteropolymeric RNA template, kim40R, that was annealed to a complementary 32-nucleotide DNA oligomer termed kim32D at a 1:4 molar ratio, as described previously (54). Reactions were conducted at 37°C with mixtures containing an RNA-DNA duplex substrate (20 nM) with RT enzymes (ϳ200 nM) in assay buffer (50 mM Tris-HCl [pH 7.8], 60 mM KCl, 5 mM MgCl 2 ) in the presence of a heparin trap (2 mg/ml).…”

Section: Chemicalsmentioning

confidence: 99%

“…In view of the high prevalence of M184I/V mutations due to the clinical use of lamivudine (3TC) and FTC, we wondered why the E138K mutation had not been seen over time in patients who failed 3TC-and/or FTC-based therapies, as might have been expected due to the compensatory effects that we and others have described (21,54). Obvious questions are whether the pressure of the M184I/V mutations might somehow prevent the emergence of the E138K mutation and whether the E138K mutation might impact the evolutionary dynamics of M184I/V-containing viruses under 3TC-FTC selection pressure.…”

mentioning

confidence: 99%

“…In addition, we recently demonstrated that recombinant HIV-1 RT enzymes containing the E138K mutation are impaired in RNase H activity and have decreased polymerization rates under conditions of high dNTP concentrations (54). 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.…”

mentioning

confidence: 99%

“…Our laboratory recently generated recombinant mutated and wild-type (WT) reverse transcriptase enzymes and HIV-1 NL4-3 infectious clones containing these mutations and demonstrated that the E138K mutation compensates for the deficit in deoxynucleoside triphosphate (dNTP) usage by the M184I/V mutations and restores both the RT enzymatic processivity and viral replication capacity of HIV-1 variants harboring the M184I/V mutations (54). The compensatory effect of the M184I/V mutations may have clinical significance in regard to treatment failures involving ETR-RPV as well as the possible presence of these mutations in transmitted resistance.…”

mentioning

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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Structure‐based design of diarylpyrimidines and triarylpyrimidines as potent HIV‐1 NNRTIs with improved metabolic stability and drug resistance profiles

Xie,

Wang,

Zhao

et al. 2024

Journal of Medical Virology

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Non‐nucleoside reverse transcriptase inhibitors (NNRTIs) are an important component of anti‐acquired immunodeficiency syndrome treatment regimen. In the present work, with the previously reported compound K‐16c as lead, a series of novel 2,4,5‐trisubstituted pyrimidine derivatives were designed based on the cocrystal structure of K‐16c/RT, with the aim to improve the anti‐human immunodeficiency virus type‐1 (HIV‐1) activities and metabolic stability properties. Compound 11b1 exhibited the most potent antiviral activity against wild‐type (WT) and a panel of single mutant HIV‐1 strains (EC50 = 2.4−12.4 nM), being superior to or comparable to those of the approved drug etravirine. Meanwhile, 11b1 exhibited moderate cytotoxicity (CC50 = 4.96 μM) and high selectivity index (SI = 1189) toward HIV‐1 WT strain. As for HIV‐1 RT inhibition test, 11b1 possessed excellent inhibitory potency (IC50 = 0.04 μM) and confirmed its target was RT. Moreover, the molecular dynamics simulation was performed to elucidate the improved drug resistance profiles. Moreover, 11b1 was demonstrated with favorable safety profiles and pharmacokinetic properties in vivo, indicating that 11b1 is a potential anti‐HIV‐1 drug candidate worthy of further development.

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Compensation by the E138K Mutation in HIV-1 Reverse Transcriptase for Deficits in Viral Replication Capacity and Enzyme Processivity Associated with the M184I/V Mutations

Cited by 83 publications

References 40 publications

Interaction of Reverse Transcriptase (RT) Mutations Conferring Resistance to Lamivudine and Etravirine: Effects on Fitness and RT Activity of Human Immunodeficiency Virus Type 1

Interaction of Reverse Transcriptase (RT) Mutations Conferring Resistance to Lamivudine and Etravirine: Effects on Fitness and RT Activity of Human Immunodeficiency Virus Type 1

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

Structure‐based design of diarylpyrimidines and triarylpyrimidines as potent HIV‐1 NNRTIs with improved metabolic stability and drug resistance profiles

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