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TitleSubstrate and target selectivity of 4'-fluoroadenosine against viral and host polymerases.
Journal, issue, pagesbioRxiv, Year 2026
Publish dateMay 26, 2026
AuthorsSimon M Walker / Arlo J Loutan / Egor P Tchesnokov / Dana Kocincova / Calvin J Gordon / Ruby A Escobedo / Nathaniel Jackson / Olivia A Vogel / Kim Morsheimer / Suncheol Park / Anant Gharpure / Ixchel Urbano / Mina Heacock / Zhong Cheng / Kushboo Pathak / Karen C Wolff / Lauren Huerta / Malina A Bakowski / Laura Riva / Anil K Gupta / Chenguang Yu / Kalyan Das / Luis Martinez-Sobrido / Christopher F Basler / Robert Davey / Ian A Wilson / Andrew B Ward / Sumit Chanda / Arnab K Chatterjee / Matthias Götte
PubMed AbstractDeveloping safe and effective treatments against emerging RNA viruses is an important goal in pandemic preparedness efforts. 4'-fluorouridine (4'-FlU) is a broad-spectrum antiviral that was shown to ...Developing safe and effective treatments against emerging RNA viruses is an important goal in pandemic preparedness efforts. 4'-fluorouridine (4'-FlU) is a broad-spectrum antiviral that was shown to inhibit viral RNA-dependent RNA polymerases (RdRps). Given its notable range of antiviral activity, this class of nucleoside analogs warrants further investigation. Here, we studied the antiviral activity and underlying mechanism of inhibition of 4'-fluoroadenosine (4'-FlA). Like 4'-FlU, 4'-FlA demonstrates a broad-spectrum of antiviral activity against eight prototypic viruses representing diverse families. Enzyme kinetics show that the triphosphate (4'-FlA-TP) is efficiently incorporated by viral RdRps. A cryo-EM structure of RdRp of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in complex with double-stranded RNA and the incorporated monophosphate (4'-FlA-MP) characterizes interactions at the active site. The incorporated analog elicits heterogeneous inhibition patterns in primer extension reactions. In contrast, templates with embedded 4'-FlA-MP inhibit incorporation of complementary UTP across the viral RdRps. However, incorporation of 4'-FIA-TP is not limited to viral polymerases and likewise includes human mitochondrial RNA polymerase. These results demonstrate the general potential for 4'-fluorinated nucleotides as antiviral drugs and guide the development of more selective derivatives for medical use in appropriate settings.
External linksbioRxiv / PubMed:42244653 / PubMed Central
MethodsEM (single particle)
Resolution3.38 Å
Structure data

76733

12sn

EMDB-76733, PDB-12sn:
SARS-CoV-2 RNA-dependent RNA polymerase in complex with 4'-FlA nucleotide analogue
Method: EM (single particle) / Resolution: 3.38 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-POP:
PYROPHOSPHATE 2-

PDB-1dcz:
BIOTIN CARBOXYL CARRIER DOMAIN OF TRANSCARBOXYLASE (TC 1.3S)

ChemComp-MG:
Unknown entry

Source
  • severe acute respiratory syndrome coronavirus 2
KeywordsVIRAL PROTEIN/DNA / Inhibitor / RNA polymerase / nucleotide analogue / SARS-CoV-2 / RdRp / VIRAL PROTEIN / VIRAL PROTEIN-DNA complex

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