RNA-activated protein cleavage with a CRISPR-associated endopeptidase
Name
nihms-1863051.pdf
Description
Accepted version
Size
2.08 MB
Format
Adobe PDF
Checksum (MD5)
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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
Date Issued
November 25, 2022
Journal
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
Abstract
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 Desulfonema ishimotonii 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.
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 Desulfonema ishimotonii 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.
MIT Department
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
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DOI of Published Version
https://doi.org/10.1126/science.add7450
