Systematic detection of poly(A)(+) RNA-interacting proteins and their differential binding

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Item Type:	Article
Title:	Systematic detection of poly(A)(+) RNA-interacting proteins and their differential binding
Creators Name:	Milek, M. and Landthaler, M.
Abstract:	RNA-binding proteins are dynamic posttranscriptional regulators of gene expression. Identification of mRNA-binding proteins in a given experimental setting is thus of great importance. We describe a procedure to enrich for direct poly(A)(+) RNA protein binders by 4-thiouridine-enhanced UV cross-linking and oligo(dT) purification. Subsequent nuclease-mediated release of RNA-binding proteins (RBPs) from mRNA allows for detection of eluted proteins by mass spectrometry. In addition, we provide a comparative approach to detect differences in RBP binding activity upon a biological stimulus.
Keywords:	Protein-RNA Interactions, RNA-Binding Proteins, Photoactivatable Ribonucleoside, 4-Thiouridine, UV Cross-Linking, Oligo(dT) Affinity Purification, Mass Spectrometry
Source:	Methods in Molecular Biology
Series Name:	Methods in Molecular Biology
Title of Book:	RNA Detection
ISSN:	1064-3745
ISBN:	978-1-4939-7212-8
Publisher:	Springer / Humana Press
Volume:	1649
Page Range:	405-417
Date:	2018
Official Publication:	https://doi.org/10.1007/978-1-4939-7213-5_27
PubMed:	View item in PubMed

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