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Structure paper

TitleCraspase is a CRISPR RNA-guided, RNA-activated protease.
Journal, issue, pagesScience, Vol. 377, Issue 6612, Page 1278-1285, Year 2022
Publish dateSep 16, 2022
AuthorsChunyi 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 AbstractThe 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 linksScience / PubMed:36007061 / PubMed Central
MethodsEM (single particle)
Resolution2.57 - 3.8 Å
Structure data

27252

8d8n

EMDB-27252, PDB-8d8n:
gRAMP non-match PFS target RNA
Method: EM (single particle) / Resolution: 3.6 Å

27257

8d97

EMDB-27257, PDB-8d97:
Apo gRAMP
Method: EM (single particle) / Resolution: 3.8 Å

27259

8d9e

EMDB-27259, PDB-8d9e:
gRAMP-match PFS target
Method: EM (single particle) / Resolution: 3.76 Å

27260

8d9f

EMDB-27260, PDB-8d9f:
gRAMP-TPR-CHAT (Craspase)
Method: EM (single particle) / Resolution: 2.71 Å

27261

8d9g

EMDB-27261, PDB-8d9g:
gRAMP-TPR-CHAT Non match PFS target RNA(Craspase)
Method: EM (single particle) / Resolution: 2.57 Å

27262

8d9h

EMDB-27262, PDB-8d9h:
gRAMP-TPR-CHAT match PFS target RNA(Craspase)
Method: EM (single particle) / Resolution: 3.6 Å

27263

8d9i

EMDB-27263, PDB-8d9i:
gRAMP non-matching PFS-with Mg
Method: EM (single particle) / Resolution: 3.62 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-PO4:
PHOSPHATE ION / Phosphate

Source
  • candidatus scalindua brodae (bacteria)
KeywordsRNA BINDING PROTEIN/RNA / CRISPR / GRAMP / RNA BINDING PROTEIN-RNA complex / RNA BINDING PROTEIN / Craspase

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