RNA-activated protein cleavage with a CRISPR-associated endopeptidase.

Autor:	Strecker J; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA., Demircioglu FE; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA., Li D; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, MA 02139, USA., Faure G; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA., Wilkinson ME; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA., Gootenberg JS; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA., Abudayyeh OO; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA., Nishimasu H; Structural Biology Division, Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo 153-8904, Japan.; Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Tokyo 113-8656, Japan.; Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo 113-0033, Japan.; Inamori Research Institute for Science, Kyoto 600-8411, Japan., Macrae RK; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA., Zhang F; Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Jazyk:	angličtina
Zdroj:	Science (New York, N.Y.) [Science] 2022 Nov 25; Vol. 378 (6622), pp. 874-881. Date of Electronic Publication: 2022 Nov 03.
DOI:	10.1126/science.add7450
Abstrakt:	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.
Databáze:	MEDLINE
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