6JDJ

Crystal structure of AcrIIC2 dimer in complex with partial Nme1Cas9

Summary for 6JDJ

• Entry DOI: 10.2210/pdb6jdj/pdb
• Descriptor: AcrIIC2, CRISPR-associated endonuclease Cas9 (3 entities in total)
• Functional Keywords: crispr-cas9, nme1cas9, nmecas9, anti-crispr, acriic2, hydrolase-hydrolase inhibitor complex, hydrolase/hydrolase inhibitor
• Biological source: Neisseria meningitidis 8013; More
• Total number of polymer chains: 3
• Total formula weight: 37184.87

Authors

Cheng, Z.,Huang, X.,Sun, W.,Wang, Y. (deposition date: 2019-02-01, release date: 2019-05-15, Last modification date: 2023-11-22)

Primary citation

Thavalingam, A.,Cheng, Z.,Garcia, B.,Huang, X.,Shah, M.,Sun, W.,Wang, M.,Harrington, L.,Hwang, S.,Hidalgo-Reyes, Y.,Sontheimer, E.J.,Doudna, J.,Davidson, A.R.,Moraes, T.F.,Wang, Y.,Maxwell, K.L.
Inhibition of CRISPR-Cas9 ribonucleoprotein complex assembly by anti-CRISPR AcrIIC2.
Nat Commun, 10:2806-2806, 2019

PubMed Abstract: CRISPR-Cas adaptive immune systems function to protect bacteria from invasion by foreign genetic elements. The CRISPR-Cas9 system has been widely adopted as a powerful genome-editing tool, and phage-encoded inhibitors, known as anti-CRISPRs, offer a means of regulating its activity. Here, we report the crystal structures of anti-CRISPR protein AcrIIC2 alone and in complex with Nme1Cas9. We demonstrate that AcrIIC2 inhibits Cas9 through interactions with the positively charged bridge helix, thereby preventing sgRNA loading. In vivo phage plaque assays and in vitro DNA cleavage assays show that AcrIIC2 mediates its activity through a large electronegative surface. This work shows that anti-CRISPR activity can be mediated through the inhibition of Cas9 complex assembly.

PubMed: 31243272
DOI: 10.1038/s41467-019-10577-3

Experimental method

X-RAY DIFFRACTION (2.6 Å)