Biochemical properties of the xenotropic murine leukemia virus-related virus integrase

Xenotropic Murine Leukemia Virus-related Virus (XMRV) is a new gammaretrovirus generated by genetic recombination between two murine endogenous retroviruses, PreXMRV1 and PreXMRV2, during passaging of human prostate cancer xenografts in laboratory mice. XMRV is representative of an early founder vir...

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Bibliographic Details
Published in:Biochimie 2014-12, Vol.107, p.300-309
Main Authors: Mbemba, Gladys, Henry, Etienne, Delelis, Olivier, Bouger, Marie-Christine, Buckle, Malcolm, Mouscadet, Jean-François, Hazan, Uriel, Leh, Hervé, Bury-Moné, Stéphanie
Format: Article
Language:English
Subjects:
3′-processing
Amino Acid Sequence
Biochemistry
Biochemistry, Molecular Biology
Desintegration
Dithiothreitol - pharmacology
Escherichia coli
Heterocyclic Compounds, 3-Ring - pharmacology
HIV Integrase - chemistry
HIV-1
Human immunodeficiency virus 1
Hydrogen-Ion Concentration
Integrase Inhibitors - pharmacology
Integrases - chemistry
Integrases - genetics
Integrases - isolation & purification
Integrases - metabolism
Life Sciences
MLV
Molecular Sequence Data
Pyrrolidinones - pharmacology
Quinolones - pharmacology
Raltegravir Potassium
Recombinant Proteins - genetics
Recombinant Proteins - isolation & purification
Retrovirus
Sequence Homology, Amino Acid
Strand transfer
Substrate Specificity
Viral Proteins - chemistry
Viral Proteins - genetics
Viral Proteins - metabolism
Xenotropic murine leukemia virus-related virus - enzymology
XMRV
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Summary:Xenotropic Murine Leukemia Virus-related Virus (XMRV) is a new gammaretrovirus generated by genetic recombination between two murine endogenous retroviruses, PreXMRV1 and PreXMRV2, during passaging of human prostate cancer xenografts in laboratory mice. XMRV is representative of an early founder virus that jumps species from mouse to human cell lines. Relatively little information is available concerning the XMRV integrase (IN), an enzyme that catalyzes a key stage in the retroviral cycle, and whose sequence is conserved among replication competent retroviruses emerging from recombination between the murine endogenous PreXMRV-1 and PreXMRV-2 genomes. Previous studies have shown that IN inhibitors efficiently block XMRV multiplication in cells. We thus aimed at characterizing the biochemical properties and sensitivity of the XMRV IN to the raltegravir, dolutegravir, 118-D-24 and elvitegravir inhibitors in vitro. We report for the first time the purification and enzymatic characterization of recombinant XMRV IN. This IN, produced in Escherichia coli and purified under native conditions, is optimally active over a pH range of 7–8.5, in the presence of Mg2+ (15 mM and 30 mM for 3′-processing and strand transfer, respectively) and is poorly sensitive to the addition of dithiothreitol. Raltegravir was shown to be a very potent inhibitor (IC50 ∼ 30 nM) whereas dolutegravir and elvitegravir were less effective (IC50 ∼ 230 nM and 650 nM, respectively). The 118-D-24 drug had no impact on XMRV IN activity. Interestingly, the substrate specificity of XMRV IN seems to be less marked compared to HIV-1 IN since XMRV IN is able to process various donor substrates that share little homology. Finally, our analysis revealed some original properties of the XMRV IN such as its relatively low sequence specificity. •First report of the purification and enzymatic characterization of the XMRV IN.•Raltegravir inhibits XMRV IN better than dolutegravir and elvitegravir.•The 118-D-24 drug had no impact on XMRV IN activity.•XMRV IN is able to process various donor substrates that share little homology.•Original properties of the XMRV IN such as its relatively low sequence specificity.
ISSN:0300-9084
1638-6183
DOI:10.1016/j.biochi.2014.09.019