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. Parniak; Kamalendra Singh; Stefan G. Sarafianos

doi:10.1093/nar/gkr694

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

Veterinary Population Medicine

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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 language	English (US)
Pages (from-to)	345-359
Number of pages	15
Journal	Nucleic acids research
Volume	40
Issue number	1
DOIs	https://doi.org/10.1093/nar/gkr694
State	Published - 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).

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Ndongwe, T. P., Adedeji, A. O., Michailidis, E., Ong, Y. T., Hachiya, A., Marchand, B., Ryan, E. M., Rai, D., Kirby, K. A., Whatley, A. S., Burke, D. H., Johnson, M., Ding, S., Zheng, Y. M., Liu, S. L., Kodama, E. I., Delviks-Frankenberry, K. A., Pathak, V. K., Mitsuya, H., ... Sarafianos, S. G. (2012). Biochemical, inhibition and inhibitor resistance studies of xenotropic murine leukemia virus-related virus reverse transcriptase. Nucleic acids research, 40(1), 345-359. https://doi.org/10.1093/nar/gkr694

Ndongwe, TP, Adedeji, AO, Michailidis, E, Ong, YT, Hachiya, A, Marchand, B, Ryan, EM, Rai, D, Kirby, KA, Whatley, AS, Burke, DH, Johnson, M, Ding, S, Zheng, YM, Liu, SL, Kodama, EI, Delviks-Frankenberry, KA, Pathak, VK, Mitsuya, H, Parniak, MA, Singh, K & Sarafianos, SG 2012, 'Biochemical, inhibition and inhibitor resistance studies of xenotropic murine leukemia virus-related virus reverse transcriptase', Nucleic acids research, vol. 40, no. 1, pp. 345-359. https://doi.org/10.1093/nar/gkr694

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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 (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.",

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

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).",

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T1 - Biochemical, inhibition and inhibitor resistance studies of xenotropic murine leukemia virus-related virus reverse transcriptase

AU - Ndongwe, Tanyaradzwa P.

AU - Adedeji, Adeyemi O.

AU - Michailidis, Eleftherios

AU - Ong, Yee Tsuey

AU - Hachiya, Atsuko

AU - Marchand, Bruno

AU - Ryan, Emily M.

AU - Rai, Devendra

AU - Kirby, Karen A.

AU - Whatley, Angela S.

AU - Burke, Donald H.

AU - Johnson, Marc

AU - Ding, Shilei

AU - Zheng, Yi Min

AU - Liu, Shan Lu

AU - Kodama, Ei Ichi

AU - Delviks-Frankenberry, Krista A.

AU - Pathak, Vinay K.

AU - Mitsuya, Hiroaki

AU - Parniak, Michael A.

AU - Singh, Kamalendra

AU - Sarafianos, Stefan G.

N1 - 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).

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N2 - 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.

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

Abstract

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