RNA-activated protein cleavage with a CRISPR-associated endopeptidase
Author(s)
Strecker, Jonathan; Demircioglu, F Esra; Li, David; Faure, Guilhem; Wilkinson, Max E; Gootenberg, Jonathan S; Abudayyeh, Omar O; Nishimasu, Hiroshi; Macrae, Rhiannon K; Zhang, Feng; ... Show more Show less
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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
<jats:italic>Desulfonema ishimotonii</jats:italic>
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.
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Date issued
2022-11-25Department
Massachusetts Institute of Technology. Department of Brain and Cognitive SciencesJournal
Science
Publisher
American Association for the Advancement of Science (AAAS)
Citation
Strecker, Jonathan, Demircioglu, F Esra, Li, David, Faure, Guilhem, Wilkinson, Max E et al. 2022. "RNA-activated protein cleavage with a CRISPR-associated endopeptidase." Science, 378 (6622).
Version: Author's final manuscript