7V94
Cryo-EM structure of the Cas12c2-sgRNA-target DNA ternary complex
Summary for 7V94
| Entry DOI | 10.2210/pdb7v94/pdb |
| EMDB information | 31807 31808 |
| Descriptor | Cas12c2, sgRNA, target DNA (target strand), ... (4 entities in total) |
| Functional Keywords | cas12c, c2c3, sgrna, target dna, crispr, rna binding protein-rna-dna complex, rna binding protein/rna/dna |
| Biological source | uncultured archaeon More |
| Total number of polymer chains | 4 |
| Total formula weight | 194795.44 |
| Authors | Kurihara, N.,Hirano, H.,Tomita, A.,Kobayashi, K.,Kusakizako, T.,Nishizawa, T.,Yamashita, K.,Nishimasu, H.,Nureki, O. (deposition date: 2021-08-24, release date: 2022-04-13, Last modification date: 2024-06-12) |
| Primary citation | Kurihara, N.,Nakagawa, R.,Hirano, H.,Okazaki, S.,Tomita, A.,Kobayashi, K.,Kusakizako, T.,Nishizawa, T.,Yamashita, K.,Scott, D.A.,Nishimasu, H.,Nureki, O. Structure of the type V-C CRISPR-Cas effector enzyme. Mol.Cell, 82:1865-1877.e4, 2022 Cited by PubMed Abstract: 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. PubMed: 35366394DOI: 10.1016/j.molcel.2022.03.006 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (2.7 Å) |
Structure validation
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