- plus-stranded RNA virus
- viral polymerase
- drug development
- enzyme inhibitor
- nucleoside/nucleotide analog
- coronavirus
- positive-sense RNA virus
- Ebola virus (EBOV)
- Middle East respiratory syndrome coronavirus (MERS–CoV)
- SARS–CoV-2
- remdesivir
- antiviral drug
- RNA chain termination
- RNA-dependent RNA polymerase (RdRp)
- viral replicase
Introduction
- Siegel D.
- Hui H.C.
- Doerffler E.
- Clarke M.O.
- Chun K.
- Zhang L.
- Neville S.
- Carra E.
- Lew W.
- Ross B.
- Wang Q.
- Wolfe L.
- Jordan R.
- Soloveva V.
- Knox J.
- et al.
- Agostini M.L.
- Andres E.L.
- Sims A.C.
- Graham R.L.
- Sheahan T.P.
- Lu X.
- Smith E.C.
- Case J.B.
- Feng J.Y.
- Jordan R.
- Ray A.S.
- Cihlar T.
- Siegel D.
- Mackman R.L.
- Clarke M.O.
- et al.
- Siegel D.
- Hui H.C.
- Doerffler E.
- Clarke M.O.
- Chun K.
- Zhang L.
- Neville S.
- Carra E.
- Lew W.
- Ross B.
- Wang Q.
- Wolfe L.
- Jordan R.
- Soloveva V.
- Knox J.
- et al.
- Agostini M.L.
- Andres E.L.
- Sims A.C.
- Graham R.L.
- Sheahan T.P.
- Lu X.
- Smith E.C.
- Case J.B.
- Feng J.Y.
- Jordan R.
- Ray A.S.
- Cihlar T.
- Siegel D.
- Mackman R.L.
- Clarke M.O.
- et al.
- Deval J.
- Hong J.
- Wang G.
- Taylor J.
- Smith L.K.
- Fung A.
- Stevens S.K.
- Liu H.
- Jin Z.
- Dyatkina N.
- Prhavc M.
- Stoycheva A.D.
- Serebryany V.
- Liu J.
- Smith D.B.
- et al.
Results
Expression of MERS–CoV RdRp complex

Inhibition of EBOV RdRp and MERS–CoV RdRp with RDV

Competitive inhibition of RNA synthesis by RDV-TP

Selectivity measurements of ATP analogs

ATP (n = 7) | RDV-TP (n = 6) | Ara-ATP (n = 5) | 2′CM-ATP (n = 5) | |
---|---|---|---|---|
Vmax (product fraction) | 0.47 ± 0.011 | 0.50 ± 0.012 | 0.48 ± 0.019 | 0.47 ± 0.021 |
% error | 2 | 2 | 4 | 4 |
Km (μm) | 0.017 ± 0.0019 | 0.0063 ± 0.00069 | 13 ± 1.7 | 2.8 ± 0.53 |
% error | 11 | 11 | 13 | 19 |
Vmax/Km | 28 | 79 | 0.037 | 0.17 |
Selectivity (fold) | 1 | 0.35 | 749 | 165 |
Discussion
- de Wilde A.H.
- Jochmans D.
- Posthuma C.C.
- Zevenhoven-Dobbe J.C.
- van Nieuwkoop S.
- Bestebroer T.M.
- van den Hoogen B.G.
- Neyts J.
- Snijder E.J.
- Arabi Y.M.
- Asiri A.Y.
- Assiri A.M.
- Aziz Jokhdar H.A.
- Alothman A.
- Balkhy H.H.
- AlJohani S.
- Al Harbi S.
- Kojan S.
- Al Jeraisy M.
- Deeb A.M.
- Memish Z.A.
- Ghazal S.
- Al Faraj S.
- Al-Hameed F.
- et al.
- Agostini M.L.
- Andres E.L.
- Sims A.C.
- Graham R.L.
- Sheahan T.P.
- Lu X.
- Smith E.C.
- Case J.B.
- Feng J.Y.
- Jordan R.
- Ray A.S.
- Cihlar T.
- Siegel D.
- Mackman R.L.
- Clarke M.O.
- et al.
- Agostini M.L.
- Andres E.L.
- Sims A.C.
- Graham R.L.
- Sheahan T.P.
- Lu X.
- Smith E.C.
- Case J.B.
- Feng J.Y.
- Jordan R.
- Ray A.S.
- Cihlar T.
- Siegel D.
- Mackman R.L.
- Clarke M.O.
- et al.
Experimental procedures
Chemicals
Protein expression and purification
RNA synthesis activity
Author contributions
Author Profile
Calvin J. Gordon
Acknowledgments
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Article info
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Footnotes
This study was supported by Canadian Institutes of Health Research Grant 159507 (to M. G.) and a grant from the Alberta Ministry of Economic Development, Trade and Tourism by the Major Innovation Fund Program for the AMR–One Health Consortium (to M. G.). M. G. has previously received funding from Gilead Sciences in support for the study of EBOV RdRp inhibition by RDV.
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