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 Embargo date 2023-07-01 ISSN 0036-8075 Source Science, 377 (6612), 1278-1285 Bibliographical note Green Open Access added to TU Delft Institutional Repository 'You share, we take care!' - Taverne project https://www.openaccess.nl/en/you-share-we-take-care Otherwise as indicated in the copyright section: the publisher is the copyright holder of this work and the author uses the Dutch legislation to make this work public. 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 science.add5064_2.pdf 2.44 MB Close viewer /islandora/object/uuid:f05e287e-4700-48e3-b88f-f7fd35a0743c/datastream/OBJ/view