{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,5,28]],"date-time":"2026-05-28T16:05:24Z","timestamp":1779984324746,"version":"3.53.1"},"posted":{"date-parts":[[2026,5,26]]},"group-title":"Microbiology","reference-count":98,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,26]],"date-time":"2026-05-26T00:00:00Z","timestamp":1779753600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"name":"CAMPP-AViDD Consortium through the National Institutes of Health","award":["1U19AI171443-01"],"award-info":[{"award-number":["1U19AI171443-01"]}]},{"name":"Alberta Ministry of Technology and Innovation through SPP-ARC"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,26]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>Developing safe and effective treatments against emerging RNA viruses is an important goal in pandemic preparedness efforts. 4\u2032-fluorouridine (4\u2032-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\u2032-fluoroadenosine (4\u2032-FlA). Like 4\u2032-FlU, 4\u2032-FlA demonstrates a broad-spectrum of antiviral activity against eight prototypic viruses representing diverse families. Enzyme kinetics show that the triphosphate (4\u2032-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\u2032-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\u2032-FlA-MP inhibit incorporation of complementary UTP across the viral RdRps. However, incorporation of 4\u2032-FIA-TP is not limited to viral polymerases and likewise includes human mitochondrial RNA polymerase. These results demonstrate the general potential for 4\u2032-fluorinated nucleotides as antiviral drugs and guide the development of more selective derivatives for medical use in appropriate settings.<\/jats:p>","DOI":"10.64898\/2026.05.22.727251","type":"posted-content","created":{"date-parts":[[2026,5,27]],"date-time":"2026-05-27T03:45:14Z","timestamp":1779853514000},"source":"Crossref","is-referenced-by-count":0,"title":["Substrate and target selectivity of 4\u2032-fluoroadenosine against viral and host polymerases"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-7231-0663","authenticated-orcid":false,"given":"Simon M.","family":"Walker","sequence":"first","affiliation":[{"name":"Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arlo J.","family":"Loutan","sequence":"additional","affiliation":[{"name":"Department of Medical Microbiology and Immunology, 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