- EMDB-34218: Structure of the Cas7-11-Csx29-guide RNA-target RNA (non-matching... -
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基本情報
登録情報
データベース: EMDB / ID: EMD-34218
タイトル
Structure of the Cas7-11-Csx29-guide RNA-target RNA (non-matching PFS) complex
マップデータ
試料
複合体: Cas7-11 in complex with a crRNA and its target RNA
複合体: crRNA
RNA: crRNA
複合体: target RNA with 3'-tag
RNA: target RNA with non-matching PFS
複合体: Cas7-11, Csx29
タンパク質・ペプチド: CRISPR-associated RAMP family protein
タンパク質・ペプチド: CHAT domain-containing protein
リガンド: CYTIDINE-5'-MONOPHOSPHATE
リガンド: ADENOSINE MONOPHOSPHATE
リガンド: ZINC ION
キーワード
CRISPR / RNase / RNA BINDING PROTEIN-RNA COMPLEX
機能・相同性
CHAT domain / CHAT domain / CRISPR type III-associated protein / RAMP superfamily / defense response to virus / CRISPR-associated RAMP family protein / CHAT domain-containing protein
Japan Agency for Medical Research and Development (AMED)
日本
引用
ジャーナル: Science / 年: 2022 タイトル: RNA-triggered protein cleavage and cell growth arrest by the type III-E CRISPR nuclease-protease. 著者: Kazuki Kato / Sae Okazaki / Cian Schmitt-Ulms / Kaiyi Jiang / Wenyuan Zhou / Junichiro Ishikawa / Yukari Isayama / Shungo Adachi / Tomohiro Nishizawa / Kira S Makarova / Eugene V Koonin / ...著者: Kazuki Kato / Sae Okazaki / Cian Schmitt-Ulms / Kaiyi Jiang / Wenyuan Zhou / Junichiro Ishikawa / Yukari Isayama / Shungo Adachi / Tomohiro Nishizawa / Kira S Makarova / Eugene V Koonin / Omar O Abudayyeh / Jonathan S Gootenberg / Hiroshi Nishimasu / 要旨: The type III-E CRISPR-Cas7-11 effector binds a CRISPR RNA (crRNA) and the putative protease Csx29 and catalyzes crRNA-guided RNA cleavage. We report cryo-electron microscopy structures of the Cas7-11- ...The type III-E CRISPR-Cas7-11 effector binds a CRISPR RNA (crRNA) and the putative protease Csx29 and catalyzes crRNA-guided RNA cleavage. We report cryo-electron microscopy structures of the Cas7-11-crRNA-Csx29 complex with and without target RNA (tgRNA), and demonstrate that tgRNA binding induces conformational changes in Csx29. Biochemical experiments revealed tgRNA-dependent cleavage of the accessory protein Csx30 by Csx29. Reconstitution of the system in bacteria showed that Csx30 cleavage yields toxic protein fragments that cause growth arrest, which is regulated by Csx31. Csx30 binds Csx31 and the associated sigma factor RpoE (RNA polymerase, extracytoplasmic E), suggesting that Csx30-mediated RpoE inhibition modulates the cellular response to infection. We engineered the Cas7-11-Csx29-Csx30 system for programmable RNA sensing in mammalian cells. Overall, the Cas7-11-Csx29 effector is an RNA-dependent nuclease-protease.