Supuni Thalalla Gamage

Abstract NAT10 is the sole eukaryotic acetyltransferase that catalyzes N4-acetylcytidine (ac4C) modification of RNA. While dysregulation of NAT10 is associated with cancer and premature aging syndromes, the requirement for its acetyltransferase activity and how NAT10 coordinates catalysis and RNA binding remain poorly understood.

MATERIALS AND METHODS Thumpd1+/− mice were generated by introduction of Cas9 protein and two synthetically modified guide RNAs (Synthego) into C57BL/6N fertilized eggs by microinjection. The synthetic guide RNAs were designed using sgRNA Scorer 2.0 (80) to target two cuts, one in exon 1 and one in exon 4. Gcn2+/− mouse was previously described (54) and obtained from the Jackson Laboratory (no. 008240). All mice were maintained and backcrossed on C57/BL6 background.

Abstract Transfer RNA (tRNA) modifications are crucial for protein synthesis, but their position-specific physiological roles remain poorly understood. Here we investigate the impact of N4-acetylcytidine (ac4C), a highly conserved tRNA modification, using a Thumpd1 knockout mouse model. We find that loss of Thumpd1-dependent tRNA acetylation leads to reduced levels of tRNALeu, increased ribosome stalling, and activation of eIF2α phosphorylation.