Some RNA and protein sequences are capable of directing changes to the course of translation from that expected from the mRNA sequence, and this process is termed translational 'recoding'. 'CHYSEL' peptides are approximately 19-amino-acid sequences found in many viral genomes. When translated at internal portions of polypeptides, they yield co-translational separation of the nascent chain at their C-termini. We dissected the reaction promoted by CHYSEL sequences using yeast genetics and in vitro translation systems. Our results indicate that the reaction occurs within the peptidyltransferase centre of the ribosome where the nascent chain is hydrolytically released from tRNA despite the presence of further sense codons.