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.
