Bulk FRET Analysis of the Conformation of the Cohesin Ring.
Cohesin is a member of the SMC protein family that exerts its chromosome functions by holding district DNA segments together using the ring structure. Topological cohesin loading involves complex, dynamic structural changes; these transitions are tightly linked to the initial cohesin association with DNA and subsequent DNA entrapment, which occurs through the transient opening of the ring at the interfaces between subunits. This chapter describes a method of monitoring the conformational changes of fission yeast cohesin that combines biochemical reconstitution and fluorescence resonance energy transfer (FRET). This method is useful for studying the structural dynamics and protein-protein interactions of not only cohesin but also other SMC proteins.
Higashi TL