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Serial interactome capture of the human cell nucleus
Please always quote using this URN: urn:nbn:de:bvb:20-opus-166172
- Novel RNA-guided cellular functions are paralleled by an increasing number of RNA-binding proteins (RBPs). Here we present ‘serial RNA interactome capture’ (serIC), a multiple purification procedure of ultraviolet-crosslinked poly(A)–RNA–protein complexes that enables global RBP detection with high specificity. We apply serIC to the nuclei of proliferating K562 cells to obtain the first human nuclear RNA interactome. The domain composition of the 382 identified nuclear RBPs markedly differs from previous IC experiments, including few factorsNovel RNA-guided cellular functions are paralleled by an increasing number of RNA-binding proteins (RBPs). Here we present ‘serial RNA interactome capture’ (serIC), a multiple purification procedure of ultraviolet-crosslinked poly(A)–RNA–protein complexes that enables global RBP detection with high specificity. We apply serIC to the nuclei of proliferating K562 cells to obtain the first human nuclear RNA interactome. The domain composition of the 382 identified nuclear RBPs markedly differs from previous IC experiments, including few factors without known RNA-binding domains that are in good agreement with computationally predicted RNA binding. serIC extends the number of DNA–RNA-binding proteins (DRBPs), and reveals a network of RBPs involved in p53 signalling and double-strand break repair. serIC is an effective tool to couple global RBP capture with additional selection or labelling steps for specific detection of highly purified RBPs.…
Author: | Thomas Conrad, Anne-Susann Albrecht, Veronica Rodrigues de Melo Costa, Sascha Sauer, David Meierhofer, Ulf Andersson Ørom |
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URN: | urn:nbn:de:bvb:20-opus-166172 |
Document Type: | Journal article |
Faculties: | Medizinische Fakultät |
Language: | English |
Parent Title (English): | Nature Communications |
Year of Completion: | 2016 |
Volume: | 7 |
Issue: | 11212 |
Source: | Nature Communications 2016, 7:11212. DOI: 10.1038/ncomms11212 |
DOI: | https://doi.org/10.1038/ncomms11212 |
Dewey Decimal Classification: | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
Tag: | RNA-binding proteins; human cell nucleus; serial RNA interactome capture |
Release Date: | 2019/07/04 |
Licence (German): | CC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International |