Elp3 uses a conserved molecular tunnel to transport acetate between distant active sites and catalyze tRNA wobble base modification.

Geissler, Evan P; Moawad, Youmna; Roehling, Paige N; Driscoll, Cassidy; Martin, Katherine; Asare-Okai, Papa Nii; Mugridge, Jeffrey S · Nat Commun · 2026

basic_science · Level V

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Abstract

The radical SAM enzyme Elp3 and eukaryotic Elongator complex catalyze formation of a key intermediate transfer RNA (tRNA) modification, 5-carboxymethyluridine (cm<sup>5</sup>U), in the anticodons of tRNAs across all domains of life. cm<sup>5</sup>U-derived modifications are important for fine tuning codon-anticodon interactions and efficient protein translation, and defects in this modification are linked to development of neurodegenerative disease in humans. Here we reconstitute tRNA modification activity with a model Elp3 enzyme and combine structural analyses, enzymology, and isotope incorporation experiments to show Elp3 harbors a conserved molecular tunnel that shuttles free acetate molecules from the acetyl-CoA binding domain to the radical SAM active site over 20 Å away, where acetate undergoes radical-mediated reaction and addition to tRNA U34. Our model explains how Elp3 bridges a large distance between active sites to catalyze tRNA carboxymethylation and illustrates a unique mechanism for intermediate transport in radical SAM enzymes.