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-Structure paper
タイトル | Cryo-EM structure and protease activity of the type III-E CRISPR-Cas effector. |
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ジャーナル・号・ページ | Nat Microbiol, Vol. 8, Issue 3, Page 522-532, Year 2023 |
掲載日 | 2023年1月26日 |
著者 | Yangao Huo / Hongshen Zhao / Qinghua Dong / Tao Jiang / |
PubMed 要旨 | The recently discovered type III-E CRISPR-Cas effector Cas7-11 shows promise when used as an RNA manipulation tool, but its structure and the mechanisms underlying its function remain unclear. Here ...The recently discovered type III-E CRISPR-Cas effector Cas7-11 shows promise when used as an RNA manipulation tool, but its structure and the mechanisms underlying its function remain unclear. Here we present four cryo-EM structures of Desulfonema ishimotonii Cas7-11-crRNA complex in pre-target and target RNA-bound states, and the cryo-EM structure of DiCas7-11-crRNA bound to its accessory protein DiCsx29. These data reveal structural elements for pre-crRNA processing, target RNA cleavage and regulation. Moreover, a 3' seed region of crRNA is involved in regulating RNA cleavage activity of DiCas7-11-crRNA-Csx29. Our analysis also shows that both the minimal mismatch of 4 nt to the 5' handle of crRNA and the minimal matching of the first 12 nt of the spacer by the target RNA are essential for triggering the protease activity of DiCas7-11-crRNA-Csx29 towards DiCsx30. Taken together, we propose that target RNA recognition and cleavage regulate and fine-tune the protease activity of DiCas7-11-crRNA-Csx29, thus preventing auto-immune responses. |
リンク | Nat Microbiol / PubMed:36702942 |
手法 | EM (単粒子) |
解像度 | 3.14 - 3.64 Å |
構造データ | EMDB-33955, PDB-7yn9: EMDB-33956, PDB-7yna: EMDB-33957, PDB-7ynb: EMDB-33958, PDB-7ync: EMDB-33959, PDB-7ynd: |
由来 |
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キーワード | RNA BINDING PROTEIN / Cas7-11 / crRNA / CRISPR-Cas / Csx29 |