Extended Data Fig. 5: Cross-linking of the Chd1 ATPase to nucleosomal DNA shows that disruption of the GSD/brace II helix allows for more robust interactions at superhelix location 2 (SHL2).
From: A competitive regulatory mechanism of the Chd1 remodeler is integral to distorting nucleosomal DNA
After labeling with the photo-crosslinker azidophenacyl bromide (APB), V721C variants of Chd1[wildtype] and Chd1[GSD/braceII]864-871-flex were incubated with dual labeled (Cy5)40-601-19(FAM) nucleosomes in different nucleotide conditions, UV irradiated, processed, and then the cleaved DNA fragments were resolved on urea denaturing gels. The cross-linking site indicated corresponds to the ATPase lobe 2 binding to SHL2 DNA, as previously reported19. These two scans show cross-linking to the two strands (and thus two sides) of the Widom 601 sequence. This gel is a representative of 3 independent experiments.
