HIV Reverse Transcriptase Fidelity, Clade Diversity, and Acquisition of Drug Resistance

Menéndez‐Arias, Luis

doi:10.1007/978-1-4614-7291-9_11

Cited by 6 publications

(8 citation statements)

References 150 publications

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“…It is well-known that lentiviral RTs (e.g. those of HIV-1, SIV or feline immunodeficiency virus) are less accurate than oncoretroviral RTs, such as those of murine leukaemia virus (MLV) or avian myeloblastosis virus5253. Our study shows that the WT HIV-2 ROD RT is only 1.6 times more faithful than the reference HIV-1 BH10 RT.…”

Section: Discussionmentioning

confidence: 65%

“…M184V) increase their DNA polymerase fidelity, there are examples of NRTI resistance mutations having little effect on the accuracy of HIV-1 RT (e.g. L74V, Q151M)5253. Our results show that in HIV-2, classwide NRTI resistance may not be associated with significant changes in the intrinsic fidelity of its RT, although a lower fitness is expected for drug-resistant viruses as a result of the lower catalytic efficiency of their RT.…”

Section: Discussionmentioning

confidence: 66%

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Fidelity of classwide-resistant HIV-2 reverse transcriptase and differential contribution of K65R to the accuracy of HIV-1 and HIV-2 reverse transcriptases

Saura

Sebastián-Martín

García‐Marquina

et al. 2017

Sci Rep

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Nucleoside reverse transcriptase (RT) inhibitors constitute the backbone of current therapies against human immunodeficiency virus type 1 and type 2 (HIV-1 and HIV-2, respectively). However, mutational pathways leading to the development of nucleoside analogue resistance are different in both types of HIV. In HIV-2, resistance to all approved nucleoside analogues is conferred by the combination of RT substitutions K65R, Q151M and M184V. Nucleotide incorporation kinetic analyses of mutant and wild-type (WT) HIV-2 RTs show that the triple-mutant has decreased catalytic efficiency due to the presence of M184V. Although similar effects were previously reported for equivalent mutations in HIV-1 RT, the HIV-2 enzymes were catalytically less efficient. Interestingly, in highly divergent HIV-1 RTs, K65R confers several-fold increased accuracy of DNA synthesis. We have determined the intrinsic fidelity of DNA synthesis of WT HIV-2 RT and mutants K65R and K65R/Q151M/M184V. Our results show that those changes in HIV-2 RT have a relatively small impact on nucleotide selectivity. Furthermore, we found that there were less than two-fold differences in error rates obtained with forward mutation assays using mutant and WT HIV-2 RTs. A different conformation of the β3-β4 hairpin loop in HIV-1 and HIV-2 RTs could probably explain the differential effects of K65R.

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Section: Discussionmentioning

confidence: 65%

Section: Discussionmentioning

confidence: 66%

Fidelity of classwide-resistant HIV-2 reverse transcriptase and differential contribution of K65R to the accuracy of HIV-1 and HIV-2 reverse transcriptases

Saura

Sebastián-Martín

García‐Marquina

et al. 2017

Sci Rep

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“…Analysis of the type of mutations produced in the shuttle vector and their sequence context can provide insight into the mechanisms underlying mutation rate variation. G→A substitutions, which were the most frequent type of change, may be in principle produced by the HIV-1 RT, which has been previously shown to exhibit a bias towards this specific substitution in some studies 21 , but not in others 22 23 . Importantly, though, G→A substitutions occurred 77% of the time (77/100 pooling env and int–vif–vpr ) at G G or G A dinucleotides, which are the canonical sequence targets of host A3G and A3D/F/H cytidine deaminases, respectively 24 , whereas the HIV-1 RT shows no such dinucleotide preferences 21 .…”

Section: Resultsmentioning

confidence: 97%

The external domains of the HIV-1 envelope are a mutational cold spot

et al. 2015

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In RNA viruses, mutations occur fast and have large fitness effects. While this affords remarkable adaptability, it can also endanger viral survival due to the accumulation of deleterious mutations. How RNA viruses reconcile these two opposed facets of mutation is still unknown. Here we show that, in human immunodeficiency virus (HIV-1), spontaneous mutations are not randomly located along the viral genome. We find that the viral mutation rate experiences a threefold reduction in the region encoding the most external domains of the viral envelope, which are strongly targeted by neutralizing antibodies. This contrasts with the hypermutation mechanisms deployed by other, more slowly mutating pathogens such as DNA viruses and bacteria, in response to immune pressure. We show that downregulation of the mutation rate in HIV-1 is exerted by the template RNA through changes in sequence context and secondary structure, which control the activity of apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3 (A3)-mediated cytidine deamination and the fidelity of the viral reverse transcriptase.

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“…On the other hand, the AffinityScript RT showed an overall error rate of 1.1 × 10 −5 at 50 °C and, in comparison with the HIV‐1 O RT, a higher tendency to generate large deletions, a characteristic also observed with wild‐type MLV RT . The error rates obtained at 50 or 55 °C for the HIV‐1 O and the AffinityScript RTs are in the low range in comparison with values reported for other retroviral RTs at 37 °C . Our fidelity estimates were obtained for DNA‐dependent DNA polymerization reactions, although studies with HIV‐1 M/B and MLV RTs have shown that overall error rates were similar or lower for RNA templates compared to DNA templates containing the same sequence .…”

Section: Discussionmentioning

confidence: 50%

“…MLV RT or AMV RT) are approximately ten times more faithful than lentiviral RTs (e.g. HIV‐1 RT) in DNA‐dependent DNA polymerization assays . Using the forward mutation assay based on the expression of lacZ , error rates of 1.4 × 10 −4 , 5.8 × 10 −5 and 1.2 × 10 −5 have been reported for wild‐type HIV‐1 M/B , HIV‐1 O and MLV RTs, respectively .…”

Section: Introductionmentioning

confidence: 99%

Temperature effects on the fidelity of a thermostable HIV‐1 reverse transcriptase

Saura

Menéndez‐Arias

2013

The FEBS Journal

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Transcriptomics and gene expression analysis are largely dependent of the availability of efficient thermostable reverse transcriptases (RTs). However, the intrinsic fidelity of DNA synthesis catalyzed by retroviral RTs is low. Reported error rates are in the range 1.2 × 10−5–6.7 × 10−4, with oncoretroviral RTs being the most faithful enzymes. Wild‐type HIV‐1 group O (HIV‐1O) RT is a thermostable polymerase that is able to synthesize cDNA at temperatures as high as 70 °C. At 37 °C, its error rate has been estimated at 5.8 × 10−5 in M13mp2 lacZ‐based forward mutation assays. However, at higher temperatures (e.g. 50 and 55 °C), the accuracy of HIV‐1O RT is increased by approximately two‐ to five‐fold. At 55 °C, the HIV‐1O RT error rate (1.3 × 10−5) was similar to that shown by the AffinityScript (Agilent Technologies Inc., La Jolla, CA, USA) RT, a commercially available thermostable murine leukaemia virus RT. At higher temperatures, the increased accuracy of the HIV‐1 enzyme results from a lower base substitution error rate, although it shows a higher tendency to introduce frameshifts. Kinetic studies carried out with model template‐primers suggest minor differences in nucleotide discrimination, although, at higher temperatures, HIV‐1O RT showed a reduced ability to extend mispaired template‐primers.

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HIV Reverse Transcriptase Fidelity, Clade Diversity, and Acquisition of Drug Resistance

Cited by 6 publications

References 150 publications

Fidelity of classwide-resistant HIV-2 reverse transcriptase and differential contribution of K65R to the accuracy of HIV-1 and HIV-2 reverse transcriptases

Fidelity of classwide-resistant HIV-2 reverse transcriptase and differential contribution of K65R to the accuracy of HIV-1 and HIV-2 reverse transcriptases

The external domains of the HIV-1 envelope are a mutational cold spot

Temperature effects on the fidelity of a thermostable HIV‐1 reverse transcriptase

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