RNA-activated protein cleavage with a CRISPR-associated endopeptidase
In prokaryotes, CRISPR-Cas systems provide adaptive immune responses against foreign genetic elements through RNA-guided nuclease activity. Recently, additional genes with non-nuclease functions have been found in genetic association with CRISPR systems, suggesting that there may be other RNA-guided non-nucleolytic enzymes. One such gene from Desulfonema ishimotonii encodes the TPR-CHAT protease Csx29, which is associated with the CRISPR effector Cas7-11. Here, we demonstrate that this CRISPR-associated protease (CASP) exhibits programmable RNA-activated endopeptidase activity against a sigma factor inhibitor to regulate a transcriptional response. Cryo-electron microscopy of an active and substrate-bound CASP complex reveals an allosteric activation mechanism that reorganizes Csx29 catalytic residues upon target RNA binding. This work reveals an RNA-guided function in nature that can be leveraged for RNA-sensing applications in vitro and in human cells.
| Errataetall: |
CommentIn: Nat Rev Genet. 2023 Feb;24(2):71. - PMID 36543986 |
|---|---|
| Medienart: |
E-Artikel |
| Erscheinungsjahr: |
2022 |
|---|---|
| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:378 |
|---|---|
| Enthalten in: |
Science (New York, N.Y.) - 378(2022), 6622 vom: 25. Nov., Seite 874-881 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Strecker, Jonathan [VerfasserIn] |
|---|
| Links: |
|---|
| 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 |
|---|
| doi: |
10.1126/science.add7450 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM349363471 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM349363471 | ||
| 003 | DE-627 | ||
| 005 | 20240108141542.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2022 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1126/science.add7450 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1248.xml |
| 035 | |a (DE-627)NLM349363471 | ||
| 035 | |a (NLM)36423276 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Strecker, Jonathan |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a RNA-activated protein cleavage with a CRISPR-associated endopeptidase |
| 264 | 1 | |c 2022 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 30.11.2022 | ||
| 500 | |a Date Revised 04.01.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a CommentIn: Nat Rev Genet. 2023 Feb;24(2):71. - PMID 36543986 | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a In prokaryotes, CRISPR-Cas systems provide adaptive immune responses against foreign genetic elements through RNA-guided nuclease activity. Recently, additional genes with non-nuclease functions have been found in genetic association with CRISPR systems, suggesting that there may be other RNA-guided non-nucleolytic enzymes. One such gene from Desulfonema ishimotonii encodes the TPR-CHAT protease Csx29, which is associated with the CRISPR effector Cas7-11. Here, we demonstrate that this CRISPR-associated protease (CASP) exhibits programmable RNA-activated endopeptidase activity against a sigma factor inhibitor to regulate a transcriptional response. Cryo-electron microscopy of an active and substrate-bound CASP complex reveals an allosteric activation mechanism that reorganizes Csx29 catalytic residues upon target RNA binding. This work reveals an RNA-guided function in nature that can be leveraged for RNA-sensing applications in vitro and in human cells | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a Endopeptidases |2 NLM | |
| 650 | 7 | |a EC 3.4.- |2 NLM | |
| 650 | 7 | |a RNA, Bacterial |2 NLM | |
| 650 | 7 | |a CRISPR-Associated Proteins |2 NLM | |
| 650 | 7 | |a RNA, Guide, CRISPR-Cas Systems |2 NLM | |
| 650 | 7 | |a Sigma Factor |2 NLM | |
| 650 | 7 | |a Bacterial Proteins |2 NLM | |
| 700 | 1 | |a Demircioglu, F Esra |e verfasserin |4 aut | |
| 700 | 1 | |a Li, David |e verfasserin |4 aut | |
| 700 | 1 | |a Faure, Guilhem |e verfasserin |4 aut | |
| 700 | 1 | |a Wilkinson, Max E |e verfasserin |4 aut | |
| 700 | 1 | |a Gootenberg, Jonathan S |e verfasserin |4 aut | |
| 700 | 1 | |a Abudayyeh, Omar O |e verfasserin |4 aut | |
| 700 | 1 | |a Nishimasu, Hiroshi |e verfasserin |4 aut | |
| 700 | 1 | |a Macrae, Rhiannon K |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Feng |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Science (New York, N.Y.) |d 1880 |g 378(2022), 6622 vom: 25. Nov., Seite 874-881 |w (DE-627)NLM000023450 |x 1095-9203 |7 nnns |
| 773 | 1 | 8 | |g volume:378 |g year:2022 |g number:6622 |g day:25 |g month:11 |g pages:874-881 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1126/science.add7450 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 378 |j 2022 |e 6622 |b 25 |c 11 |h 874-881 | ||