Limited evidence for protein products of non-coding transcripts in the HEK293T cellular cytosol
| Autor: | Annelies Bogaert, Daria Fijalkowska, An Staes, Tessa Van de Steene, Hans Demol, Kris Gevaert |
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| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: |
TRANSLATION INITIATION LANDSCAPE
RNA Untranslated IDENTIFICATION Biology and Life Sciences PEPTIDES N-TERMINAL PROTEOMICS Biochemistry SMALL ORFS ANNOTATION Analytical Chemistry RNAS Open Reading Frames Cytosol HEK293 Cells MAMMALIAN-CELLS Protein Biosynthesis DISCOVERY REVEALS Medicine and Health Sciences Humans Peptides Ribosomes Molecular Biology |
| Zdroj: | MOLECULAR & CELLULAR PROTEOMICS |
| ISSN: | 1535-9476 1535-9484 |
| Popis: | Ribosome profiling has revealed translation outside canonical coding sequences, including translation of short upstream ORFs, long noncoding RNAs, overlapping ORFs, ORFs in UTRs, or ORFs in alternative reading frames. Studies combining mass spectrometry, ribosome profiling, and CRISPR-based screens showed that hundreds of ORFs derived from noncoding transcripts produce (micro)proteins, whereas other studies failed to find evidence for such types of noncanonical translation products. Here, we attempted to discover translation products from noncoding regions by strongly reducing the complexity of the sample prior to mass spectrometric analysis. We used an extended database as the search space and applied stringent filtering of the identified peptides to find evidence for novel translation events. We show that, theoretically our strategy facilitates the detection of translation events of transcripts from non -coding regions but experimentally only find 19 peptides that might originate from such translation events. Finally, Virotrap-based interactome analysis of two N-terminal proteoforms originating from noncoding regions showed the functional potential of these novel proteins. |
| Databáze: | OpenAIRE |
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