In vitro analysis of ribonucleoprotein complex remodeling and disassembly

Abstract

Ribonucleoprotein (RNP) complexes play essential roles in gene expression. Their assembly and disassembly control the fate of mRNA molecules. Here, we describe a method that examines the remodeling and disassembly of RNPs. One unique aspect of this method is that the RNA-binding proteins (RBPs) of interest are produced in HeLa cells with or without the desired modification and the RNP is assembled in cellular extracts with synthetic RNA oligonucleotides. We use this method to investigate how ubiquitination of an RBP affects its ability to bind its RNA target.