Desthiobiotin (DTB)-modified and TAMRA-modified 2″OMeNAD⁺ are RNA 2'-phosphotransferase (Tpt1) poisons that enable affinity-tagging and fluorescence-tagging of internal RNA 2'-phosphate groups.

ABSTRACT: 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. We showed previously that 2″OMeNAD⁺, a synthetic NAD⁺ analog that cannot support step 2 transesterification, is an effective step 1 substrate for Runella slithyformis Tpt1 (RslTpt1) in a reaction that generates the normally undetectable RNA-2'-phospho-(ADP-ribose) intermediate as an abortive product. Here we report the chemical synthesis of two novel 2″OMeNAD⁺ compounds, co

SUBMITTER: Kasprzyk R 

PROVIDER: S-EPMC12621583 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature