Structural and functional characterization of plant ARGONAUTE MID domains

Abstract

The interaction of small silencing RNA 5′ nucleotides with the MID domain of ARGONAUTE (AGO) proteins provides an anchor point that contributes to strong binding between RNA and protein. The following protocols describe the necessary procedures to characterize the structure of AGO MID domains using X-ray crystallography as well as their interaction with nucleotides that mimic the 5′ end of small silencing RNAs using two-dimensional NMR spectroscopy.