期刊
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
卷 142, 期 3, 页码 1348-1358出版社
AMER CHEMICAL SOC
DOI: 10.1021/jacs.9b10521
关键词
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资金
- Brown University
- COBRE Center for Computational Biology of Human Disease (NIGMS) [P20GM109035]
- National Science Foundation [CHE-1905374]
- NIH [R01 EY027440]
- XSEDE [TG-MCB160059]
CRISPR-Cas9 is a widely employed genome-editing tool with functionality reliant on the ability of the Cas9 endonuclease to introduce site-specific breaks in double-stranded DNA. In this system, an intriguing allosteric communication has been suggested to control its DNA cleavage activity through flexibility of the catalytic HNH domain. Here, solution NMR experiments and a novel Gaussian-accelerated molecular dynamics (GaMD) simulation method are used to capture the structural and dynamic determinants of allosteric signaling within the HNH domain. We reveal the existence of a millisecond time scale dynamic pathway that spans HNH from the region interfacing the adjacent RuvC nuclease and propagates up to the DNA recognition lobe in full-length CRISPR-Cas9. These findings reveal a potential route of signal transduction within the CRISPR-Cas9 HNH nuclease, advancing our understanding of the allosteric pathway of activation. Further, considering the role of allosteric signaling in the specificity of CRISPR-Cas9, this work poses the mechanistic basis for novel engineering efforts aimed at improving its genome-editing capability.
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