Structural basis for the promiscuous PAM recognition by Corynebacterium diphtheriae Cas9

Seiichi Hirano, Omar O. Abudayyeh, Jonathan S. Gootenberg, Takuro Horii, Ryuichiro Ishitani, Izuho Hatada, Feng Zhang, Hiroshi Nishimasu () and Osamu Nureki ()
Additional contact information
Seiichi Hirano: Graduate School of Science, The University of Tokyo
Omar O. Abudayyeh: Broad Institute of MIT and Harvard
Jonathan S. Gootenberg: Broad Institute of MIT and Harvard
Takuro Horii: Gunma University
Ryuichiro Ishitani: Graduate School of Science, The University of Tokyo
Izuho Hatada: Gunma University
Feng Zhang: Broad Institute of MIT and Harvard
Hiroshi Nishimasu: Graduate School of Science, The University of Tokyo
Osamu Nureki: Graduate School of Science, The University of Tokyo

Nature Communications, 2019, vol. 10, issue 1, 1-11

Abstract: 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.

Date: 2019
References: Add references at CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/s41467-019-09741-6 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-09741-6

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-019-09741-6

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().