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-Structure paper
タイトル | RNA targeting and cleavage by the type III-Dv CRISPR effector complex. |
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ジャーナル・号・ページ | Nat Commun, Vol. 15, Issue 1, Page 3324, Year 2024 |
掲載日 | 2024年4月18日 |
著者 | Evan A Schwartz / Jack P K Bravo / Mohd Ahsan / Luis A Macias / Caitlyn L McCafferty / Tyler L Dangerfield / Jada N Walker / Jennifer S Brodbelt / Giulia Palermo / Peter C Fineran / Robert D Fagerlund / David W Taylor / |
PubMed 要旨 | CRISPR-Cas are adaptive immune systems in bacteria and archaea that utilize CRISPR RNA-guided surveillance complexes to target complementary RNA or DNA for destruction. Target RNA cleavage at regular ...CRISPR-Cas are adaptive immune systems in bacteria and archaea that utilize CRISPR RNA-guided surveillance complexes to target complementary RNA or DNA for destruction. Target RNA cleavage at regular intervals is characteristic of type III effector complexes. Here, we determine the structures of the Synechocystis type III-Dv complex, an apparent evolutionary intermediate from multi-protein to single-protein type III effectors, in pre- and post-cleavage states. The structures show how multi-subunit fusion proteins in the effector are tethered together in an unusual arrangement to assemble into an active and programmable RNA endonuclease and how the effector utilizes a distinct mechanism for target RNA seeding from other type III effectors. Using structural, biochemical, and quantum/classical molecular dynamics simulation, we study the structure and dynamics of the three catalytic sites, where a 2'-OH of the ribose on the target RNA acts as a nucleophile for in line self-cleavage of the upstream scissile phosphate. Strikingly, the arrangement at the catalytic residues of most type III complexes resembles the active site of ribozymes, including the hammerhead, pistol, and Varkud satellite ribozymes. Our work provides detailed molecular insight into the mechanisms of RNA targeting and cleavage by an important intermediate in the evolution of type III effector complexes. |
リンク | Nat Commun / PubMed:38637512 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 2.5 - 3.44 Å |
構造データ | EMDB-40248, PDB-8s9t: EMDB-40250, PDB-8s9v: EMDB-40251, PDB-8s9x: EMDB-40276: CRISPR-Cas type III-D effector complex consensus map EMDB-40296: CRISPR-Cas type III-D effector complex local refinement map EMDB-40297: CRISPR-Cas type III-D effector complex bound to a target RNA local refinement map EMDB-40298: CRISPR-Cas type III-D effector complex bound to a target RNA consensus map |
化合物 | ChemComp-MG: ChemComp-HOH: |
由来 |
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キーワード | RNA BINDING PROTEIN / CRISPR / CRISPR-Cas / type III / complex / RNA / crRNA / RNA BINDING PROTEIN/RNA / RNA BINDING PROTEIN-RNA complex |