Summary information and primary citation

PDB-id
5f9r; SNAP-derived features in text and JSON formats; DNAproDB
Class
hydrolase-DNA-RNA
Method
X-ray (3.4 Å)
Summary
Crystal structure of catalytically-active streptococcus pyogenes crispr-cas9 in complex with single-guided RNA and double-stranded DNA primed for target DNA cleavage
Reference
Jiang F, Taylor DW, Chen JS, Kornfeld JE, Zhou K, Thompson AJ, Nogales E, Doudna JA (2016): "Structures of a CRISPR-Cas9 R-loop complex primed for DNA cleavage." Science, 351, 867-871. doi: 10.1126/science.aad8282.
Abstract
Bacterial adaptive immunity and genome engineering involving the CRISPR (clustered regularly interspaced short palindromic repeats)-associated (Cas) protein Cas9 begin with RNA-guided DNA unwinding to form an RNA-DNA hybrid and a displaced DNA strand inside the protein. The role of this R-loop structure in positioning each DNA strand for cleavage by the two Cas9 nuclease domains is unknown. We determine molecular structures of the catalytically active Streptococcus pyogenes Cas9 R-loop that show the displaced DNA strand located near the RuvC nuclease domain active site. These protein-DNA interactions, in turn, position the HNH nuclease domain adjacent to the target DNA strand cleavage site in a conformation essential for concerted DNA cutting. Cas9 bends the DNA helix by 30°, providing the structural distortion needed for R-loop formation.

Cartoon-block schematics in six views (download the tarball)

PyMOL session file Download PDB file View in 3Dmol.js