Two-step mechanism and step-arrest mutants of Runella slithyformis NAD⁺-dependent tRNA 2'-phosphotransferase Tpt1.

ABSTRACT: Tpt1 catalyzes the transfer of an internal 2'-monophosphate moiety (2'-PO₄) from a "branched" 2'-PO₄ RNA splice junction to NAD⁺ to form a "clean" 2'-OH, 3'-5' phosphodiester junction, ADP-ribose 1″-2″ cyclic phosphate, and nicotinamide. First discovered as an essential component of the Saccharomyces cerevisiae tRNA splicing machinery, Tpt1 is widely distributed in nature, including in taxa that have no yeast-like RNA splicing system. Here we characterize the RslTpt1 protein from the bacterium Runella slithyformis, in which Tpt1 is encoded within a putative RNA repair gene cluster. We find that (i) expression of RslTpt1 in yeast complements a lethal tpt1Δ knockout, and (ii) purified recombinant RslTpt1 is a bona fide NAD⁺-dep

SUBMITTER: Munir A 

PROVIDER: S-EPMC6097658 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature