6O56

HNH Nuclease from S. pyogenes Cas9

Summary for 6O56

• Entry DOI: 10.2210/pdb6o56/pdb
• Descriptor: CRISPR-associated endonuclease Cas9/Csn1 (2 entities in total)
• Functional Keywords: nuclease, crispr cas9, dna binding protein
• Biological source: Streptococcus pyogenes serotype M1
• Total number of polymer chains: 2
• Total formula weight: 31970.09

Authors

Newton, J.C.,Lisi, G.P.,Jogl, G. (deposition date: 2019-03-01, release date: 2020-01-15, Last modification date: 2023-10-11)

Primary citation

East, K.W.,Newton, J.C.,Morzan, U.N.,Narkhede, Y.B.,Acharya, A.,Skeens, E.,Jogl, G.,Batista, V.S.,Palermo, G.,Lisi, G.P.
Allosteric Motions of the CRISPR-Cas9 HNH Nuclease Probed by NMR and Molecular Dynamics.
J.Am.Chem.Soc., 142:1348-1358, 2020

PubMed Abstract: 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.

PubMed: 31885264
DOI: 10.1021/jacs.9b10521

Experimental method

X-RAY DIFFRACTION (1.9 Å)