+Search query
-Structure paper
Title | Craspase is a CRISPR RNA-guided, RNA-activated protease. |
---|---|
Journal, issue, pages | Science, Vol. 377, Issue 6612, Page 1278-1285, Year 2022 |
Publish date | Sep 16, 2022 |
Authors | Chunyi Hu / Sam P B van Beljouw / Ki Hyun Nam / Gabriel Schuler / Fran Ding / Yanru Cui / Alicia Rodríguez-Molina / Anna C Haagsma / Menno Valk / Martin Pabst / Stan J J Brouns / Ailong Ke / |
PubMed Abstract | The CRISPR-Cas type III-E RNA-targeting effector complex gRAMP/Cas7-11 is associated with a caspase-like protein (TPR-CHAT/Csx29) to form Craspase (CRISPR-guided caspase). Here, we use cryo-electron ...The CRISPR-Cas type III-E RNA-targeting effector complex gRAMP/Cas7-11 is associated with a caspase-like protein (TPR-CHAT/Csx29) to form Craspase (CRISPR-guided caspase). Here, we use cryo-electron microscopy snapshots of Craspase to explain its target RNA cleavage and protease activation mechanisms. Target-guide pairing extending into the 5' region of the guide RNA displaces a gating loop in gRAMP, which triggers an extensive conformational relay that allosterically aligns the protease catalytic dyad and opens an amino acid side-chain-binding pocket. We further define Csx30 as the endogenous protein substrate that is site-specifically proteolyzed by RNA-activated Craspase. This protease activity is switched off by target RNA cleavage by gRAMP and is not activated by RNA targets containing a matching protospacer flanking sequence. We thus conclude that Craspase is a target RNA-activated protease with self-regulatory capacity. |
External links | Science / PubMed:36007061 / PubMed Central |
Methods | EM (single particle) |
Resolution | 2.57 - 3.8 Å |
Structure data | EMDB-27252, PDB-8d8n: EMDB-27257, PDB-8d97: EMDB-27259, PDB-8d9e: EMDB-27260, PDB-8d9f: EMDB-27261, PDB-8d9g: EMDB-27262, PDB-8d9h: EMDB-27263, PDB-8d9i: |
Chemicals | ChemComp-ZN: ChemComp-PO4: |
Source |
|
Keywords | RNA BINDING PROTEIN/RNA / CRISPR / GRAMP / RNA BINDING PROTEIN-RNA complex / RNA BINDING PROTEIN / Craspase |