Study of translational control using cell-free translation systems and primer extension inhibition assays

This work examines translational regulation using fungal cell‑free translation systems and primer extension inhibition assays. Results show that the Neurospora crassa arginine attenuator peptide (AAP) can transiently stall ribosomes in multiple contexts, with stalling occurring while the AAP remains in peptidyl‑tRNA form within the exit tunnel. The foot‑and‑mouth disease virus 2A peptide similarly induces a ribosome pause that promotes peptidyl‑tRNA hydrolysis. Additionally, the Salmonella protein FljA inhibits fliC translation by binding the 5′‑UTR and blocking ribosome access. These findings demonstrate the utility of in‑vitro assays for dissecting mechanisms of translational control.