{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T18:19:31Z","timestamp":1788459571203,"version":"build-2803163510"},"reference-count":0,"publisher":"Cold Spring Harbor Laboratory","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["RNA"],"accepted":{"date-parts":[[2026,8,25]]},"abstract":"<jats:p>\n                    RNA 2'-phosphotransferase Tpt1 catalyzes the removal of an internal RNA 2'-PO4 via a two-step mechanism in which: (i) the 2'-PO4 attacks NAD+ C1'' to form an RNA-2'-phospho-(ADP-ribose) intermediate and nicotinamide; and (ii) transesterification of the ADP-ribose O2'' to the RNA 2'-phosphodiester yields 2'-OH RNA and ADP-ribose-1'',2''-cyclic phosphate. Although Tpt1 enzymes are prevalent in bacteria, archaea, and eukarya, Tpt1 is uniquely essential in fungi, where it erases the 2'-PO4 mark installed by tRNA ligases during tRNA splicing. A Tpt1 \u201cpoison\u201d that arrests the reaction after step 1 could aid in the design of antifungals that interdict tRNA biogenesis. Whereas our previous studies of 2''OMeNAD+ established its efficacy in poisoning Runella slithyformis Tpt1 (RslTpt1, a bacterial enzyme), its application was limited insofar as Tpt1 enzymes from fungal pathogens were unable to utilize 2''OMeNAD\n                    <jats:sup>+<\/jats:sup>\n                    for step 1 catalysis. Here we report the chemical synthesis of 2''F-NAD\n                    <jats:sup>+<\/jats:sup>\n                    , which proved to be a broad-spectrum poison against Tpt1 enzymes from five different taxa, including human pathogens Candida albicans and Candida auris. The resulting RNA-2'-phospho-(2''F-ADP-ribose) dead-end product remains stably trapped in a complex with Tpt1. A modified analog, 2''F-NAD-DTB, containing desthiobiotin (DTB) linked to adenine C2, provides an improved affinity-tag probe of RNA 2'-phosphate modification.\n                  <\/jats:p>","DOI":"10.1261\/rna.081192.126","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T17:45:16Z","timestamp":1788457516000},"page":"rna.081192.126","source":"Crossref","is-referenced-by-count":0,"title":["2''F-NAD\n                    <sup>+<\/sup>\n                    is a broad-spectrum RNA 2'-phosphotransferase (Tpt1) poison that traps the enzyme on its abortive RNA-2'-phospho-(2''F-ADP-ribose) reaction intermediate"],"prefix":"10.1261","author":[{"given":"Renata","family":"Kasprzyk","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shreya","family":"Ghosh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5034-6438","authenticated-orcid":false,"given":"Stewart","family":"Shuman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"246","published-online":{"date-parts":[[2026,9,3]]},"container-title":["RNA"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1261\/rna.081192.126","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T17:45:16Z","timestamp":1788457516000},"score":1,"resource":{"primary":{"URL":"http:\/\/rnajournal.cshlp.org\/lookup\/doi\/10.1261\/rna.081192.126"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,3]]},"references-count":0,"alternative-id":["10.1261\/rna.081192.126"],"URL":"https:\/\/doi.org\/10.1261\/rna.081192.126","relation":{},"ISSN":["1355-8382","1469-9001"],"issn-type":[{"value":"1355-8382","type":"print"},{"value":"1469-9001","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,3]]}}}