In vivo cross-linking followed by PolyA enrichment to identify yeast mRNA binding proteins

Abstract

mRNA binding proteins regulate gene expression by controlling the processing, localization, decay, and translation of messenger RNAs (mRNAs). To fully understand this process, it is necessary to identify the complete set of mRNA binding proteins. This work describes a method for the systematic identifi cation of yeast mRNA binding proteins. This method applies in vivo UV cross-linking, affi nity pull-down of polyA(+) mRNAs, and analysis by mass spectrometry to identify proteins that directly bind to mRNAs.