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

定量质谱和 PAR-CLIP 鉴定 RNA-蛋白质相互作用

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作者:Marion Scheibe, Falk Butter, Markus Hafner, Thomas Tuschl, Matthias Mann

Abstract

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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