title: Biochemical properties of the xenotropic murine leukemia virus-related virus integrase.
abstract: 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 Mg(2+) (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.
authors.affiliation: LBPA, CNRS, Ecole Normale Supérieure (ENS) de Cachan, 61 avenue du Président Wilson, 94235 Cachan, France. Electronic address: gladys.mbemba@lbpa.ens-cachan.fr.
authors.forename: Gladys
authors.initials: G
authors.lastname: Mbemba
authors.affiliation: LBPA, CNRS, Ecole Normale Supérieure (ENS) de Cachan, 61 avenue du Président Wilson, 94235 Cachan, France. Electronic address: ehenry@lbpa.ens-chachan.fr.
authors.forename: Etienne
authors.initials: E
authors.lastname: Henry
authors.affiliation: LBPA, CNRS, Ecole Normale Supérieure (ENS) de Cachan, 61 avenue du Président Wilson, 94235 Cachan, France. Electronic address: delelis@lbpa.ens-cachan.fr.
authors.forename: Olivier
authors.initials: O
authors.lastname: Delelis
authors.affiliation: LBPA, CNRS, Ecole Normale Supérieure (ENS) de Cachan, 61 avenue du Président Wilson, 94235 Cachan, France. Electronic address: bouger@ens-cachan.fr.
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Bouger
authors.affiliation: LBPA, CNRS, Ecole Normale Supérieure (ENS) de Cachan, 61 avenue du Président Wilson, 94235 Cachan, France. Electronic address: buckle@lbpa.ens-cachan.fr.
authors.forename: Malcolm
authors.initials: M
authors.lastname: Buckle
authors.affiliation: LBPA, CNRS, Ecole Normale Supérieure (ENS) de Cachan, 61 avenue du Président Wilson, 94235 Cachan, France; Bio-Rad Laboratories, 3 Bvd Raymond Poincaré, 92430 Marnes-la-Coquette, France(1). Electronic address: Jean-francois_Mouscadet@bio-rad.com.
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Mouscadet
authors.affiliation: LBPA, CNRS, Ecole Normale Supérieure (ENS) de Cachan, 61 avenue du Président Wilson, 94235 Cachan, France. Electronic address: uriel.hazan@ens-cachan.fr.
authors.forename: Uriel
authors.initials: U
authors.lastname: Hazan
authors.affiliation: LBPA, CNRS, Ecole Normale Supérieure (ENS) de Cachan, 61 avenue du Président Wilson, 94235 Cachan, France. Electronic address: leh@lbpa.ens-cachan.fr.
authors.forename: Hervé
authors.initials: H
authors.lastname: Leh
authors.affiliation: LBPA, CNRS, Ecole Normale Supérieure (ENS) de Cachan, 61 avenue du Président Wilson, 94235 Cachan, France. Electronic address: stephanie.bury-mone@lbpa.ens-cachan.fr.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Bury-Moné
mesh_terms: D000595:Amino Acid Sequence; D004229:Dithiothreitol / Q000494:pharmacology; D019427:HIV Integrase / Q000737:chemistry; D006575:Heterocyclic Compounds, 3-Ring / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D019429:Integrase Inhibitors / Q000494:pharmacology*; D019426:Integrases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D010078:Oxazines; D010879:Piperazines; D011728:Pyridones; D011760:Pyrrolidinones / Q000494:pharmacology; D015363:Quinolones / Q000494:pharmacology; D000068898:Raltegravir Potassium; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D058008:Xenotropic murine leukemia virus-related virus / Q000201:enzymology*; D000099226:Dolutegravir
chemical_list: D004229:Dithiothreitol; D019427:HIV Integrase; D006575:Heterocyclic Compounds, 3-Ring; D019429:Integrase Inhibitors; D019426:Integrases; D010078:Oxazines; D010879:Piperazines; D011728:Pyridones; D011760:Pyrrolidinones; D015363:Quinolones; D000068898:Raltegravir Potassium; D011994:Recombinant Proteins; D014764:Viral Proteins; D000099226:Dolutegravir; C509700:elvitegravir; C514801:p31 integrase protein, Human immunodeficiency virus 1
keywords: 3′-processing; Desintegration; HIV-1; MLV; Strand transfer; XMRV