Print Email Facebook Twitter Inhibition of SARS-CoV-2 polymerase by nucleotide analogs from a single-molecule perspective Title Inhibition of SARS-CoV-2 polymerase by nucleotide analogs from a single-molecule perspective Author Seifert, Mona (Friedrich-Alexander-Universität Erlangen-Nürnberg) Bera, Subhas C. (Friedrich-Alexander-Universität Erlangen-Nürnberg) van Nies, Pauline (Friedrich-Alexander-Universität Erlangen-Nürnberg) Kirchdoerfer, Robert N. (University of Wisconsin-Madison) Shannon, Ashleigh (Aix Marseille Université) Le, Thi Tuyet Nhung (Aix Marseille Université) Meng, Xiangzhi (University of Texas Health Science Center at San Antonio) Depken, S.M. (TU Delft BN/Bionanoscience; Kavli institute of nanoscience Delft) Dulin, David (Friedrich-Alexander-Universität Erlangen-Nürnberg; Vrije Universiteit Amsterdam) Department BN/Bionanoscience Date 2021 Abstract The absence of 'shovel-ready' anti-coronavirus drugs during vaccine development has exceedingly worsened the SARS-CoV-2 pandemic. Furthermore, new vaccine-resistant variants and coronavirus outbreaks may occur in the near future, and we must be ready to face this possibility. However, efficient antiviral drugs are still lacking to this day, due to our poor understanding of the mode of incorporation and mechanism of action of nucleotides analogs that target the coronavirus polymerase to impair its essential activity. Here, we characterize the impact of remdesivir (RDV, the only FDA-approved anti-coronavirus drug) and other nucleotide analogs (NAs) on RNA synthesis by the coronavirus polymerase using a high-throughput, single-molecule, magnetic-tweezers platform. We reveal that the location of the modification in the ribose or in the base dictates the catalytic pathway(s) used for its incorporation. We show that RDV incorporation does not terminate viral RNA synthesis, but leads the polymerase into backtrack as far as 30 nt, which may appear as termination in traditional ensemble assays. SARS-CoV-2 is able to evade the endogenously synthesized product of the viperin antiviral protein, ddhCTP, though the polymerase incorporates this NA well. This experimental paradigm is essential to the discovery and development of therapeutics targeting viral polymerases. Subject antiviral drugshigh throughput magnetic tweezersinfectious diseasemechanism of actionmicrobiologyphysics of living systemsRemdesivirSARS-CoV-2single molecule biophysicsvirus To reference this document use: http://resolver.tudelft.nl/uuid:f2aa172c-fbd7-4abf-a9e8-6be772058c98 DOI https://doi.org/10.7554/eLife.70968 ISSN 2050-084X Source eLife, 10 Part of collection Institutional Repository Document type journal article Rights © 2021 Mona Seifert, Subhas C. Bera, Pauline van Nies, Robert N. Kirchdoerfer, Ashleigh Shannon, Thi Tuyet Nhung Le, Xiangzhi Meng, S.M. Depken, David Dulin, More Authors Files PDF elife_70968_v1.pdf 9.08 MB Close viewer /islandora/object/uuid:f2aa172c-fbd7-4abf-a9e8-6be772058c98/datastream/OBJ/view