Evolutionary remodeling of non-canonical ORF translation in mammals
Chang, Y.; Lei, T.; Zhou, F.; Jiang, J.; Huang, Y.; Zhu, Z.; Zhang, H.
10.1101/2025.09.16.676541 bioRxivShow abstract
Non-canonical open reading frames (ncORFs) are pervasive within transcripts annotated as "non-coding" or "untranslated regions" of mRNAs, yet their landscape under normal physiological conditions remains to be fully resolved, particularly outside humans. Here we applied a stringent and standardized pipeline to hundreds of high-quality ribosome profiling libraries from normal mammalian tissues and cell types, identifying 11,623 human and 16,485 mouse ncORFs. Evolutionary analyses revealed that thousands of ncORFs are subject to coding constraint and exhibit lineage-specific conservation, underscoring their functional potential. Ancient ncORFs are preferentially highly translated, broadly expressed, and enriched for lineage-specific conservation. Co-expression patterns further indicate that many ncORFs, especially ancient ones, are cotranslated with canonical coding sequences, consistent with functions mediated through protein-protein interactions. Together, these findings establish a comprehensive atlas of mammalian ncORFs and provide fundamental insights into their evolutionary dynamics and functional integration within the proteome.
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