Identification of Non-Canonical Translation Products in C. elegans Using Tandem Mass Spectrometry.

Transcriptome and ribosome sequencing have revealed the existence of many non-canonical transcripts, mainly containing splice variants, ncRNA, sORFs and altORFs. However, identification and characterization of products that may be translated out of these remains a challenge. Addressing this, we here report on 552 non-canonical proteins and splice variants in the model organism C. elegans using tandem mass spectrometry . Aided by sequencing-based prediction, we generated a custom proteome database tailored to search for non-canonical translation products of C. elegans . Using this database, we mined available mass spectrometric resources of C. elegans , from which 51 novel, non-canonical proteins could be identified . Furthermore, we utilized diverse proteomic and peptidomic strategies to detect 40 novel non-canonical proteins in C. elegans by LC-TIMS-MS/MS, of which 6 were common with our meta-analysis of existing resources. Together, this permits us to provide a resource with detailed annotation of 467 splice variants and 85 novel proteins mapped onto UTRs, non-coding regions and alternative open reading frames of the C. elegans genome.
Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
(Copyright © 2021 Parmar, Peeters, Boonen, Clark, Baggerman, Menschaert and Temmerman.)