Figure 2

The MTA family subunits bind directly the H3 tail peptide. A. the H3 tail peptide-binding specificity of individual subunits of NURD complex. Each subunit of NURD complex was synthesized and ³⁵S-met labeled via in vitro coupled transcription and translation. The proteins were then subjected to in vitro pulldown with immobilized H3 tail peptides as indicated. The binding was visualized by autoradiography. Note that the MTA1 family proteins, CHD3 and RbAp48 bound H3 tail in a H3K4me3-sensitive manner, whereas p66/68 and HDAC1 were not. B. UV-induced cross-linking demonstrated a direct association of MTA family proteins with H3 tail peptide. The H3(Bpa) peptide contained at K9 position a Bpa moiety that mediated a UV-induced cross-linking with associated protein(s) in close proximity. Note that once cross-linked to the H3(Bpa) peptide, the associated protein(s) became resistant to wash with 0.2% SDS buffer.