PDBsum entry 6mcc

Hydrolase/RNA/viral protein

PDB id: 6mcc

Contents

Protein chains

1364 a.a.

130 a.a.

DNA/RNA

PDB id:

6mcc

Name:

Hydrolase/RNA/viral protein

Title:

Cryoem structure of acriia2 homolog in complex with crispr-cas9

Structure:

Single guide RNA (116-mer). Chain: b. Engineered: yes. Crispr-associated endonuclease cas9. Chain: a. Engineered: yes. Anti-crispr acriia2 homolog. Chain: c. Synonym: acriia2b.3.

Source:

Streptococcus pyogenes m1 gas. Organism_taxid: 160490. Expressed in: escherichia coli. Expression_system_taxid: 562. Gene: cas9, m1gas476_0830. Listeria phage lp-101. Organism_taxid: 1458856. Expression_system_taxid: 562

Authors:

F.Jiang,J.J.Liu,J.A.Doudna

Key ref:

F.Jiang et al. (2019). Temperature-Responsive Competitive Inhibition of CRISPR-Cas9. Mol Cell, 73, 601. PubMed id: 30595438 DOI: 10.1016/j.molcel.2018.11.016

Date:

31-Aug-18 Release date: 16-Jan-19

Protein chain

No UniProt id for this chain

Struc: 1364 a.a.

Protein chain

A0A4V8H027  (A0A4V8H027_9CAUD) -  Anti-CRISPR AcrIIA2 Homolog from Listeria phage LP-101

Seq: 130 a.a.

Struc: 130 a.a.

DNA/RNA chain

GTP-G-C-G-C-A-U-A-A-A-G-A-U-G-A-G-A-C-G-C-G-U-U-U-U-A-G-A-G-C-U-A-U-G-C-U-G-U- 116 bases

Enzyme reactions

• Enzyme class: Chain A: E.C.3.1.-.-

DOI no: 10.1016/j.molcel.2018.11.016

Mol Cell 73:601 (2019)

PubMed id: 30595438

Temperature-Responsive Competitive Inhibition of CRISPR-Cas9.

F.Jiang, J.J.Liu, B.A.Osuna, M.Xu, J.D.Berry, B.J.Rauch, E.Nogales, J.Bondy-Denomy, J.A.Doudna.

ABSTRACT

CRISPR-Cas immune systems utilize RNA-guided nucleases to protect bacteria from bacteriophage infection. Bacteriophages have in turn evolved inhibitory "anti-CRISPR" (Acr) proteins, including six inhibitors (AcrIIA1-AcrIIA6) that can block DNA cutting and genome editing by type II-A CRISPR-Cas9 enzymes. We show here that AcrIIA2 and its more potent homolog, AcrIIA2b, prevent Cas9 binding to DNA by occluding protein residues required for DNA binding. Cryo-EM-determined structures of AcrIIA2 or AcrIIA2b bound to S. pyogenes Cas9 reveal a mode of competitive inhibition of DNA binding that is distinct from other known Acrs. Differences in the temperature dependence of Cas9 inhibition by AcrIIA2 and AcrIIA2b arise from differences in both inhibitor structure and the local inhibitor-binding environment on Cas9. These findings expand the natural toolbox for regulating CRISPR-Cas9 genome editing temporally, spatially, and conditionally.