Summary information and primary citation
- PDB-id
- 6joo; SNAP-derived features in text and JSON formats;
DNAproDB
- Class
- hydrolase-DNA-RNA
- Method
- X-ray (2.9 Å)
- Summary
- Crystal structure of corynebacterium diphtheriae cas9 in complex with sgrna and target DNA
- Reference
- Hirano S, Abudayyeh OO, Gootenberg JS, Horii T, Ishitani R, Hatada I, Zhang F, Nishimasu H, Nureki O (2019): "Structural basis for the promiscuous PAM recognition by Corynebacterium diphtheriae Cas9." Nat Commun, 10, 1968. doi: 10.1038/s41467-019-09741-6.
- Abstract
- The RNA-guided DNA endonuclease Cas9 cleaves double-stranded DNA targets bearing a protospacer adjacent motif (PAM) and complementarity to an RNA guide. Unlike other Cas9 orthologs, Corynebacterium diphtheriae Cas9 (CdCas9) recognizes the promiscuous NNRHHHY PAM. However, the CdCas9-mediated PAM recognition mechanism remains unknown. Here, we report the crystal structure of CdCas9 in complex with the guide RNA and its target DNA at 2.9 Å resolution. The structure reveals that CdCas9 recognizes the NNRHHHY PAM via a combination of van der Waals interactions and base-specific hydrogen bonds. Moreover, we find that CdCas9 exhibits robust DNA cleavage activity with the optimal 22-nucleotide length guide RNAs. Our findings highlight the mechanistic diversity of the PAM recognition by Cas9 orthologs, and provide a basis for the further engineering of the CRISPR-Cas9 genome-editor nucleases.
