In:
PLOS Genetics, Public Library of Science (PLoS), Vol. 19, No. 9 ( 2023-9-20), p. e1010930-
Abstract:
Cas12g is an endonuclease belonging to the type V RNA-guided CRISPR–Cas family. It is known for its ability to cleave RNA substrates using a conserved endonuclease active site located in the RuvC domain. In this study, we determined the crystal structure of apo-Cas12g, the cryo-EM structure of the Cas12g-sgRNA binary complex and investigated conformational changes that occur during the transition from the apo state to the Cas12g-sgRNA binary complex. The conserved zinc finger motifs in Cas12g undergo an ordered-to-disordered transition from the apo to the sgRNA-bound state and their mutations negatively impact on target RNA cleavage. Moreover, we identified a lid motif in the RuvC domain that undergoes transformation from a helix to loop to regulate the access to the RuvC active site and subsequent cleavage of the RNA substrate. Overall, our study provides valuable insights into the mechanisms by which Cas12g recognizes sgRNA and the conformational changes it undergoes from sgRNA binding to the activation of the RNase active site, thereby laying a foundation for the potential repurposing of Cas12g as a tool for RNA-editing.
Type of Medium:
Online Resource
ISSN:
1553-7404
DOI:
10.1371/journal.pgen.1010930
DOI:
10.1371/journal.pgen.1010930.g001
DOI:
10.1371/journal.pgen.1010930.g002
DOI:
10.1371/journal.pgen.1010930.g003
DOI:
10.1371/journal.pgen.1010930.g004
DOI:
10.1371/journal.pgen.1010930.g005
DOI:
10.1371/journal.pgen.1010930.g006
DOI:
10.1371/journal.pgen.1010930.s001
DOI:
10.1371/journal.pgen.1010930.s002
DOI:
10.1371/journal.pgen.1010930.s003
DOI:
10.1371/journal.pgen.1010930.s004
DOI:
10.1371/journal.pgen.1010930.s005
DOI:
10.1371/journal.pgen.1010930.s006
DOI:
10.1371/journal.pgen.1010930.s007
DOI:
10.1371/journal.pgen.1010930.s008
DOI:
10.1371/journal.pgen.1010930.s009
DOI:
10.1371/journal.pgen.1010930.s010
DOI:
10.1371/journal.pgen.1010930.s011
DOI:
10.1371/journal.pgen.1010930.s012
DOI:
10.1371/journal.pgen.1010930.s013
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2023
detail.hit.zdb_id:
2186725-2
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