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Recent technological advances have expanded the annotated protein coding content of mammalian genomes, as hundreds of previously unidentified, short open reading frame (ORF)-encoded peptides (SEPs) have now been found to be translated. Although several studies have identified important physiological roles for this emerging protein class, a general method to define their interactomes is lacking. Here, we demonstrate that genetic incorporation of the photo-crosslinking noncanonical amino acid AbK into SEP transgenes allows for the facile identification of SEP cellular interaction partners using affinity-based methods. From a survey of seven SEPs, we report the discovery of short ORF-encoded histone binding protein (SEHBP), a conserved microprotein that interacts with chromatin-associated proteins, localizes to discrete genomic loci, and induces a robust transcriptional program when overexpressed in human cells. This work affords a straightforward method to help define the physiological roles of SEPs and demonstrates its utility by identifying SEHBP as a short ORF-encoded transcription factor.


Minseob Koh, Insha Ahmad, Yeonjin Ko, Yuxiang Zhang, Thomas F Martinez, Jolene K Diedrich, Qian Chu, James J Moresco, Michael A Erb, Alan Saghatelian, Peter G Schultz, Michael J Bollong. A short ORF-encoded transcriptional regulator. Proceedings of the National Academy of Sciences of the United States of America. 2021 Jan 26;118(4)

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PMID: 33468658

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