RNA-triggered protein cleavage and cell growth arrest by the type III-E CRISPR nuclease-protease
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.
| Errataetall: |
CommentIn: Nat Rev Genet. 2023 Feb;24(2):71. - PMID 36543986 |
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| Medienart: |
E-Artikel |
| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:378 |
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| Enthalten in: |
Science (New York, N.Y.) - 378(2022), 6622 vom: 25. Nov., Seite 882-889 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Kato, Kazuki [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 30.11.2022 Date Revised 04.01.2024 published: Print-Electronic CommentIn: Nat Rev Genet. 2023 Feb;24(2):71. - PMID 36543986 Citation Status MEDLINE |
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| doi: |
10.1126/science.add7347 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 500 | |a CommentIn: Nat Rev Genet. 2023 Feb;24(2):71. - PMID 36543986 | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a 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 | ||
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| 650 | 4 | |a Research Support, N.I.H., Extramural | |
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| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
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| 700 | 1 | |a Okazaki, Sae |e verfasserin |4 aut | |
| 700 | 1 | |a Schmitt-Ulms, Cian |e verfasserin |4 aut | |
| 700 | 1 | |a Jiang, Kaiyi |e verfasserin |4 aut | |
| 700 | 1 | |a Zhou, Wenyuan |e verfasserin |4 aut | |
| 700 | 1 | |a Ishikawa, Junichiro |e verfasserin |4 aut | |
| 700 | 1 | |a Isayama, Yukari |e verfasserin |4 aut | |
| 700 | 1 | |a Adachi, Shungo |e verfasserin |4 aut | |
| 700 | 1 | |a Nishizawa, Tomohiro |e verfasserin |4 aut | |
| 700 | 1 | |a Makarova, Kira S |e verfasserin |4 aut | |
| 700 | 1 | |a Koonin, Eugene V |e verfasserin |4 aut | |
| 700 | 1 | |a Abudayyeh, Omar O |e verfasserin |4 aut | |
| 700 | 1 | |a Gootenberg, Jonathan S |e verfasserin |4 aut | |
| 700 | 1 | |a Nishimasu, Hiroshi |e verfasserin |4 aut | |
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