Print Email Facebook Twitter Craspase is a CRISPR RNA-guided, RNA-activated protease Title Craspase is a CRISPR RNA-guided, RNA-activated protease Author Hu, Chunyi (Cornell University) van Beljouw, S.P.B. (TU Delft BN/Stan Brouns Lab; Kavli institute of nanoscience Delft) Nam, Ki Hyun (Pohang University of Science and Technology) Schuler, Gabriel (Cornell University) Rodríguez Molina, A. (TU Delft BN/Stan Brouns Lab; Kavli institute of nanoscience Delft) van Eijkeren-Haagsma, A.C. (TU Delft BN/Stan Brouns Lab; Kavli institute of nanoscience Delft) Valk, M. (TU Delft BN/Stan Brouns Lab; Kavli institute of nanoscience Delft) Pabst, Martin (TU Delft BT/Environmental Biotechnology) Brouns, S.J.J. (TU Delft BN/Stan Brouns Lab; Kavli institute of nanoscience Delft) Date 2022 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 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. To reference this document use: http://resolver.tudelft.nl/uuid:f05e287e-4700-48e3-b88f-f7fd35a0743c DOI https://doi.org/10.1126/science.add5064 ISSN 0036-8075 Source Science, 377 (6612), 1278-1285 Bibliographical note Accepted Author Manuscript Part of collection Institutional Repository Document type journal article Rights © 2022 Chunyi Hu, S.P.B. van Beljouw, Ki Hyun Nam, Gabriel Schuler, A. Rodríguez Molina, A.C. van Eijkeren-Haagsma, M. Valk, Martin Pabst, S.J.J. Brouns, More Authors Files PDF Hu_2022_Science_repository_002_.pdf 5.63 MB Close viewer /islandora/object/uuid:f05e287e-4700-48e3-b88f-f7fd35a0743c/datastream/OBJ/view