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Quantitative mass spectrometry and PAR-CLIP to identify RNA-protein interactions

Systematic analysis of the RNA-protein interactome requires robust and scalable methods. We here show the combination of two completely orthogonal, generic techniques to identify RNA-protein interactions: PAR-CLIP reveals a collection of RNAs bound to a protein whereas SILAC-based RNA pull-downs ide...

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Detalles Bibliográficos
Autores principales: Scheibe, Marion, Butter, Falk, Hafner, Markus, Tuschl, Thomas, Mann, Matthias
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2012
Materias:
RNA
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3479200/
https://www.ncbi.nlm.nih.gov/pubmed/22885304
http://dx.doi.org/10.1093/nar/gks746
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author Scheibe, Marion
Butter, Falk
Hafner, Markus
Tuschl, Thomas
Mann, Matthias
author_facet Scheibe, Marion
Butter, Falk
Hafner, Markus
Tuschl, Thomas
Mann, Matthias
author_sort Scheibe, Marion
collection PubMed
description Systematic analysis of the RNA-protein interactome requires robust and scalable methods. We here show the combination of two completely orthogonal, generic techniques to identify RNA-protein interactions: PAR-CLIP reveals a collection of RNAs bound to a protein whereas SILAC-based RNA pull-downs identify a group of proteins bound to an RNA. We investigated binding sites for five different proteins (IGF2BP1-3, QKI and PUM2) exhibiting different binding patterns. We report near perfect agreement between the two approaches. Nevertheless, they are non-redundant, and ideally complement each other to map the RNA-protein interaction network.
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spelling pubmed-34792002012-10-24 Quantitative mass spectrometry and PAR-CLIP to identify RNA-protein interactions Scheibe, Marion Butter, Falk Hafner, Markus Tuschl, Thomas Mann, Matthias Nucleic Acids Res RNA Systematic analysis of the RNA-protein interactome requires robust and scalable methods. We here show the combination of two completely orthogonal, generic techniques to identify RNA-protein interactions: PAR-CLIP reveals a collection of RNAs bound to a protein whereas SILAC-based RNA pull-downs identify a group of proteins bound to an RNA. We investigated binding sites for five different proteins (IGF2BP1-3, QKI and PUM2) exhibiting different binding patterns. We report near perfect agreement between the two approaches. Nevertheless, they are non-redundant, and ideally complement each other to map the RNA-protein interaction network. Oxford University Press 2012-10 2012-08-09 /pmc/articles/PMC3479200/ /pubmed/22885304 http://dx.doi.org/10.1093/nar/gks746 Text en © The Author(s) 2012. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle RNA
Scheibe, Marion
Butter, Falk
Hafner, Markus
Tuschl, Thomas
Mann, Matthias
Quantitative mass spectrometry and PAR-CLIP to identify RNA-protein interactions
title Quantitative mass spectrometry and PAR-CLIP to identify RNA-protein interactions
title_full Quantitative mass spectrometry and PAR-CLIP to identify RNA-protein interactions
title_fullStr Quantitative mass spectrometry and PAR-CLIP to identify RNA-protein interactions
title_full_unstemmed Quantitative mass spectrometry and PAR-CLIP to identify RNA-protein interactions
title_short Quantitative mass spectrometry and PAR-CLIP to identify RNA-protein interactions
title_sort quantitative mass spectrometry and par-clip to identify rna-protein interactions
topic RNA
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3479200/
https://www.ncbi.nlm.nih.gov/pubmed/22885304
http://dx.doi.org/10.1093/nar/gks746
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