Fig. 3: Structures of Cas9 spanning the entire reaction from a single dataset.
From: Visualization of a multi-turnover Cas9 after product release
a Close-up view of the intact scissile bond of the target strand (TS) in states I and II. Map density shown from state II. b Close-up view of the cleaved scissile bond showing clear separation between nucleotides 3 and 4 of the hybridized TS. Map density shown from state III. c Close-up view of the cut site in state V following PAM-distal product release. PAM-proximal product remains bound where the first three nucleotides of the TS upstream of the PAM remain hybridized. d The five main conformational states populate the entire reaction cycle where (I) represents the pre-activation state just following R-loop completion, (II) represents the pre-cleavage state during reorganization of the NUC lobe on-path to dock at the cleavage sites, (III) represents the conformational checkpoint state where HNH undocks following DNA cleavage, (IV) represents the product state where HNH remains flexibly tethered after undocking from the cut TS, and (V) represents the dissociated state where the PAM-distal end of the DNA is released from the complex.
