Structure of the type V-C CRISPR-Cas effector enzyme.

Bacterial Proteins / metabolism CRISPR-Associated Proteins / metabolism CRISPR-Cas Systems Cryoelectron Microscopy DNA / genetics RNA, Guide, Kinetoplastida / metabolism Ribonucleases / metabolism
CRISPR Cas12c cryo-EM

Journal

Molecular cell
ISSN: 1097-4164
Titre abrégé: Mol Cell
Pays: United States
ID NLM: 9802571

Informations de publication

Date de publication:
19 05 2022
Historique:
received: 28 08 2021
revised: 27 12 2021
accepted: 28 02 2022
pubmed: 3 4 2022
medline: 25 5 2022
entrez: 2 4 2022
Statut: ppublish

Résumé

RNA-guided CRISPR-Cas nucleases are widely used as versatile genome-engineering tools. Recent studies identified functionally divergent type V Cas12 family enzymes. Among them, Cas12c2 binds a CRISPR RNA (crRNA) and a trans-activating crRNA (tracrRNA) and recognizes double-stranded DNA targets with a short TN PAM. Here, we report the cryo-electron microscopy structures of the Cas12c2-guide RNA binary complex and the Cas12c2-guide RNA-target DNA ternary complex. The structures revealed that the crRNA and tracrRNA form an unexpected X-junction architecture, and that Cas12c2 recognizes a single T nucleotide in the PAM through specific hydrogen-bonding interactions with two arginine residues. Furthermore, our biochemical analyses indicated that Cas12c2 processes its precursor crRNA to a mature crRNA using the RuvC catalytic site through a unique mechanism. Collectively, our findings improve the mechanistic understanding of diverse type V CRISPR-Cas effectors.

Identifiants

DOI: 10.1016/j.molcel.2022.03.006 PMID: 35366394 PMC: PMC9522604
pubmed: 35366394
pii: S1097-2765(22)00215-5
doi: 10.1016/j.molcel.2022.03.006
pmc: PMC9522604
mid: EMS156764
pii:
doi:

Substances chimiques

Bacterial Proteins 0
CRISPR-Associated Proteins 0
RNA, Guide 0
DNA 9007-49-2
Ribonucleases EC 3.1.-

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

1865-1877.e4

Subventions

Organisme : Medical Research Council
ID : MC_UP_A025_1012
Pays : United Kingdom

Informations de copyright

Copyright © 2022 Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of interests O.N. is a co-founder, board member, and scientific advisor for Modalis and Curreio.

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Auteurs

Nina Kurihara (N)

Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.

Ryoya Nakagawa (R)

Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.

Hisato Hirano (H)

Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.

Sae Okazaki (S)

Structural Biology Division, Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.

Atsuhiro Tomita (A)

Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.

Kan Kobayashi (K)

PeptiDream Inc., 3-25-23 Tonomachi, Kawasaki-ku, Kawasaki City, Kanagawa Prefecture 210-0821, Japan.

Tsukasa Kusakizako (T)

Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.

Tomohiro Nishizawa (T)

Graduate School of Medical Life Science, Yokohama City University, Yokohama 230-0045, Japan.

Keitaro Yamashita (K)

MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.

David A Scott (DA)

Arbor Biotechnologies, Cambridge, MA 02139, USA.

Hiroshi Nishimasu (H)

Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan; Structural Biology Division, Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan; Inamori Research Institute for Science, 620 Suiginya-cho, Shimogyo-ku, Kyoto 600-8411, Japan. Electronic address: nisimasu@g.ecc.u-tokyo.ac.jp.

Osamu Nureki (O)

Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. Electronic address: nureki@bs.s.u-tokyo.ac.jp.

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Classifications MeSH

questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines bactériennes › Structure of the type V-C CRISPR-Cas effector enzyme.
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines associées aux CRISPR › Structure of the type V-C CRISPR-Cas effector enzyme.
questionsmedicales.fr › Phénomènes génétiques › Régulation de l'expression des gènes › Épigenèse génétique › Extinction de l'expression des gènes › Systèmes CRISPR-Cas › Structure of the type V-C CRISPR-Cas effector enzyme.
questionsmedicales.fr › Techniques d'investigation › Microscopie électronique › Cryomicroscopie électronique › Structure of the type V-C CRISPR-Cas effector enzyme.
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ADN › Structure of the type V-C CRISPR-Cas effector enzyme.
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ARN › ARN non traduit › Petit ARN non traduit › ARN guide › Structure of the type V-C CRISPR-Cas effector enzyme.
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Hydrolases › Esterases › Ribonucléases › Structure of the type V-C CRISPR-Cas effector enzyme.