biostudies-literatureElder JJHNational Institute of Environmental Health SciencesIntramural NIH HHSNational Institutes of HealthNational Institute of General Medical SciencesNIGMS NIH HHS511-525https://www.ebi.ac.uk/biostudies/studies/S-EPMC11152995biostudies-literatureUnknown40(6)Ribonucleases (RNases) play important roles in supporting canonical and non-canonical roles of tRNAs by catalyzing the cleavage of the tRNA phosphodiester backbone. Here, we highlight how recent advances in cryo-electron microscopy (cryo-EM), protein structure prediction, reconstitution experiments, tRNA sequencing, and other studies have revealed new insight into the nucleases that process tRNA. This represents a very diverse group of nucleases that utilize distinct mechanisms to recognize and cleave tRNA during different stages of a tRNA's life cycle including biogenesis, fragmentation, surveillance, and decay. In this review, we provide a synthesis of the structure, mechanism, regulation, and modes of tRNA recognition by tRNA nucleases, along with open questions for future investigationTrends in genetics : TIGThe making and breaking of tRNAs by ribonucleases.PMC11152995ZIA ES103247R00 GM143534R00GM143534Stanley REHayne CKPapadopoulos RElder JJHfalseThe making and breaking of tRNAs by ribonucleases.Ribonucleases (RNases) play important roles in supporting canonical and non-canonical roles of tRNAs by catalyzing the cleavage of the tRNA phosphodiester backbone. Here, we highlight how recent advances in cryo-electron microscopy (cryo-EM), protein structure prediction, reconstitution experiments, tRNA sequencing, and other studies have revealed new insight into the nucleases that process tRNA. This represents a very diverse group of nucleases that utilize distinct mechanisms to recognize and cleave tRNA during different stages of a tRNA's life cycle including biogenesis, fragmentation, surveillance, and decay. In this review, we provide a synthesis of the structure, mechanism, regulation, and modes of tRNA recognition by tRNA nucleases, along with open questions for future investigation2024-01-01T00:00:00Z2024 Jun2026-06-03T07:33:30.919Z2026-04-26T03:09:46.515ZS-EPMC111529953864147110.1016/j.tig.2024.03.007