Author/Editor		Milek, Miha; Landthaler, Markus
Title		Systematic Detection of Poly(A)+ RNA-Interacting Proteins and Their Differential Binding
Type		članek
Source		In: RNA detection : methods and protocols New York, NY : Humana Press
Publication year		2018
Volume		str. 405-417
ISSN		1064-3745 - Methods in molecular biology (Clifton, N.J.)
Language		eng
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		interakcije beljakovin-RNA RNA-vezavni proteini masna spektrometrija protein-RNA interactions RNA-binding proteins mass spectrometry