Biochemical, inhibition and inhibitor resistance studies of xenotropic murine leukemia virus-related virus reverse transcriptase

Tanyaradzwa P. Ndongwe, Adeyemi O. Adedeji, Eleftherios Michailidis, Yee Tsuey Ong, Atsuko Hachiya, Bruno Marchand, Emily M. Ryan, Devendra Rai, Karen A. Kirby, Angela S. Whatley, Donald H. Burke, Marc Johnson, Shilei Ding, Yi Min Zheng, Shan Lu Liu, Ei Ichi Kodama, Krista A. Delviks-Frankenberry, Vinay K. Pathak, Hiroaki Mitsuya, Michael A. ParniakKamalendra Singh, Stefan G. Sarafianos

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

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 (koff) 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.

Original languageEnglish (US)
Pages (from-to)345-359
Number of pages15
JournalNucleic acids research
Volume40
Issue number1
DOIs
StatePublished - Jan 2012

Bibliographical note

Funding Information:
NIH grants (AI076119, AI079801, and AI094715, to S.G.S.), (AI074389, to D.H.B.), (AI079801 to M.A.P.); NIH Bench-to-Bedside Award and the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research (to V.K.P); Ministry of Knowledge and Economy, Bilateral International Collaborative R&D Program, Republic of Korea; Canadian Institutes of Health Research (CIHR) and University of Missouri (to S-L.L.); amfAR Mathilde Krim Fellowship and a CIHR Fellowship (to B.M.). Funding for open access charge: NIH grants (AI076119, AI094715, AI074389, AI079801).

Fingerprint

Dive into the research topics of 'Biochemical, inhibition and inhibitor resistance studies of xenotropic murine leukemia virus-related virus reverse transcriptase'. Together they form a unique fingerprint.

Cite this