An In Vitro Assay to Study RSC Remodeling Activity Stimulation by the Cohesin Loader

The cohesin loader complex, consisting in Scc2 and Scc4 subunits, allows cohesin access to chromosomes, and therefore, it is required for cohesin's essential roles in many structural and functional aspects of chromosomes such as sister chromatid cohesion, genome organization, gene transcription, and DNA repair. The cohesin loader interacts with the RSC chromatin remodeling complex directing cohesin loading to nucleosome-free regions. In addition to stimulate cohesin loading and define the cohesin loading sites along the genome, the cohesin loader plays a role in nucleosome positioning at gene promoters, suggesting an interplay between cohesin loading and chromatin remodeling. Here, we describe an in vitro assay to assess the ability of the cohesin loader to stimulate RSC nucleosome sliding activity.