The human RNA-binding proteome and its dynamics during translational arrest
Summary - Proteins and RNA functionally and physically intersect in multiple biological processes, however, currently no universal method is available to purify protein-RNA complexes. Here, we introduce XRNAX, a method for the generic purification of protein-crosslinked RNA, and demonstrate its vers...
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| Main Authors: | , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
2019
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| In: |
Cell
Year: 2018, Volume: 176, Issue: 1, Pages: 391-403 |
| ISSN: | 1097-4172 |
| DOI: | 10.1016/j.cell.2018.11.004 |
| Online Access: | Verlag, Volltext: https://doi.org/10.1016/j.cell.2018.11.004 Verlag, Volltext: http://www.sciencedirect.com/science/article/pii/S0092867418314636 
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| Author Notes: | Jakob Trendel, Thomas Schwarzl, Rastislav Horos, Ananth Prakash, Alex Bateman, Matthias W. Hentze, and Jeroen Krijgsveld |
| Summary: | Summary - Proteins and RNA functionally and physically intersect in multiple biological processes, however, currently no universal method is available to purify protein-RNA complexes. Here, we introduce XRNAX, a method for the generic purification of protein-crosslinked RNA, and demonstrate its versatility to study the composition and dynamics of protein-RNA interactions by various transcriptomic and proteomic approaches. We show that XRNAX captures all RNA biotypes and use this to characterize the sub-proteomes that interact with coding and non-coding RNAs (ncRNAs) and to identify hundreds of protein-RNA interfaces. Exploiting the quantitative nature of XRNAX, we observe drastic remodeling of the RNA-bound proteome during arsenite-induced stress, distinct from autophagy-related changes in the total proteome. In addition, we combine XRNAX with crosslinking immunoprecipitation sequencing (CLIP-seq) to validate the interaction of ncRNA with lamin B1 and EXOSC2. Thus, XRNAX is a resourceful approach to study structural and compositional aspects of protein-RNA interactions to address fundamental questions in RNA-biology. |
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| Item Description: | Published: December 6, 2018 Gesehen am 03.05.2019 |
| Physical Description: | Online Resource |
| ISSN: | 1097-4172 |
| DOI: | 10.1016/j.cell.2018.11.004 |