Ultraviolet Photochemical Cross-Linking to Detect RNA-Binding Proteins

Abstract

RNA is photoreactive on exposure to ultraviolet (UV) light in the 250- to 270-nm range. On UV treatment, RNA bases absorb energy to generate free radicals that can covalently attach to nearby amino acid residues in RNA-bound proteins. UV cross-linking experiments have been extensively used to identify and characterize RNA-binding proteins. The method described here involves the use of 32P-labeled RNA and crude extracts or purified proteins.