title: Biochemical, inhibition and inhibitor resistance studies of xenotropic murine leukemia virus-related virus reverse transcriptase.
abstract: We report key mechanistic differences between the reverse transcriptases (RT) of human immunodeficiency virus type-1 (HIV-1) and of xenotropic murine leukemia virus-related virus (XMRV), a gammaretrovirus that can infect human cells. Steady and pre-steady state kinetics demonstrated that XMRV RT is significantly less efficient in DNA synthesis and in unblocking chain-terminated primers. Surface plasmon resonance experiments showed that the gammaretroviral enzyme has a remarkably higher dissociation rate (k(off)) from DNA, which also results in lower processivity than HIV-1 RT. Transient kinetics of mismatch incorporation revealed that XMRV RT has higher fidelity than HIV-1 RT. We identified RNA aptamers that potently inhibit XMRV, but not HIV-1 RT. XMRV RT is highly susceptible to some nucleoside RT inhibitors, including Translocation Deficient RT inhibitors, but not to non-nucleoside RT inhibitors. We demonstrated that XMRV RT mutants K103R and Q190M, which are equivalent to HIV-1 mutants that are resistant to tenofovir (K65R) and AZT (Q151M), are also resistant to the respective drugs, suggesting that XMRV can acquire resistance to these compounds through the decreased incorporation mechanism reported in HIV-1.
authors: Christopher Bond Life Sciences Center, Department of Molecular Microbiology & Immunology, University of Missouri, School of Medicine, Columbia, MO 65211, USA.
authors: Tanyaradzwa P
authors: TP
authors: Ndongwe
authors: Adeyemi O
authors: AO
authors: Adedeji
authors: Eleftherios
authors: E
authors: Michailidis
authors: Yee Tsuey
authors: YT
authors: Ong
authors: Atsuko
authors: A
authors: Hachiya
authors: Bruno
authors: B
authors: Marchand
authors: Emily M
authors: EM
authors: Ryan
authors: Devendra K
authors: DK
authors: Rai
authors: Karen A
authors: KA
authors: Kirby
authors: Angela S
authors: AS
authors: Whatley
authors: Donald H
authors: DH
authors: Burke
authors: Marc
authors: M
authors: Johnson
authors: Shilei
authors: S
authors: Ding
authors: Yi-Min
authors: YM
authors: Zheng
authors: Shan-Lu
authors: SL
authors: Liu
authors: Ei-Ichi
authors: E
authors: Kodama
authors: Krista A
authors: KA
authors: Delviks-Frankenberry
authors: Vinay K
authors: VK
authors: Pathak
authors: Hiroaki
authors: H
authors: Mitsuya
authors: Michael A
authors: MA
authors: Parniak
authors: Kamalendra
authors: K
authors: Singh
authors: Stefan G
authors: SG
authors: Sarafianos
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000595:Amino Acid Sequence; D052157:Aptamers, Nucleotide / Q000494:pharmacology; D004247:DNA / Q000096:biosynthesis / Q000378:metabolism; D054303:HIV Reverse Transcriptase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000201:enzymology; D009154:Mutation; D009711:Nucleotides / Q000378:metabolism; D063065:Organophosphonates / Q000494:pharmacology; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D000068698:Tenofovir; D058008:Xenotropic murine leukemia virus-related virus / Q000201:enzymology*; D015215:Zidovudine / Q000494:pharmacology; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D052157:Aptamers, Nucleotide; D009711:Nucleotides; D063065:Organophosphonates; D018894:Reverse Transcriptase Inhibitors; D015215:Zidovudine; D004247:DNA; D000068698:Tenofovir; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase; D001616:beta-Galactosidase; D000225:Adenine