Enhanced super-resolution ribosome profiling unveils pervasive translation of upstream ORFs and small ORFs in Arabidopsis

In recent years, the discovery that the translational regulation of many mRNAs is influenced by small upstream open reading frames (uORFs) has highlighted the importance of precisely identifying gene structures and translated ORFs to understand gene functions and cellular behavior. Using Arabidopsis, Wu et al. advanced the precision of ribosome profiling, in which RNA sequences bound by ribosomes are identified, revealing 7,751 unconventional translation occurrences. Subsequent validation via proteomic scrutiny affirmed the production of stable proteins from the identified sites. Noteworthy instances of active translation were observed in small interfering RNAs and microRNAs. A novel methodology was developed to discern short upstream ORFs (uORFs), showcasing distinct translational repression patterns. Moreover, the recognition of 594 uORFs regulated through alternative splicing, and their involvement in crucial pathways, underscores the comprehensive nature of this investigation in elucidating the mechanisms governing gene expression regulation in Arabidopsis. (Summary by Yueh Cho @YuehCho1984) Plant Cell. 10.1093/plcell/koad290.