IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v10y2019i1d10.1038_s41467-019-09741-6.html
   My bibliography  Save this article

Structural basis for the promiscuous PAM recognition by Corynebacterium diphtheriae Cas9

Author

Listed:
  • Seiichi Hirano

    (Graduate School of Science, The University of Tokyo)

  • Omar O. Abudayyeh

    (Broad Institute of MIT and Harvard
    Massachusetts Institute of Technology)

  • Jonathan S. Gootenberg

    (Broad Institute of MIT and Harvard
    Massachusetts Institute of Technology)

  • 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
    Massachusetts Institute of Technology
    Massachusetts Institute of Technology
    Massachusetts Institute of Technology)

  • Hiroshi Nishimasu

    (Graduate School of Science, The University of Tokyo)

  • Osamu Nureki

    (Graduate School of Science, The University of Tokyo)

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.

Suggested Citation

  • Seiichi Hirano & Omar O. Abudayyeh & Jonathan S. Gootenberg & Takuro Horii & Ryuichiro Ishitani & Izuho Hatada & Feng Zhang & Hiroshi Nishimasu & Osamu Nureki, 2019. "Structural basis for the promiscuous PAM recognition by Corynebacterium diphtheriae Cas9," Nature Communications, Nature, vol. 10(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-09741-6
    DOI: 10.1038/s41467-019-09741-6
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-019-09741-6
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/s41467-019-09741-6?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-09741-6. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    We have no bibliographic references for this item. You can help adding them by using this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.