7KIX
The structure of anti-CRISPR AcrIE2
Summary for 7KIX
| Entry DOI | 10.2210/pdb7kix/pdb |
| NMR Information | BMRB: 30807 |
| Descriptor | Anti-CRISPR protein AcrIE2 (1 entity in total) |
| Functional Keywords | viral protein |
| Biological source | Pseudomonas phage JBD16C |
| Total number of polymer chains | 1 |
| Total formula weight | 10319.22 |
| Authors | Pawluk, A.,Davidson, A.R.,Maxwell, K.L. (deposition date: 2020-10-25, release date: 2021-05-26, Last modification date: 2024-05-01) |
| Primary citation | Mejdani, M.,Pawluk, A.,Maxwell, K.L.,Davidson, A.R. Anti-CRISPR AcrIE2 Binds the Type I-E CRISPR-Cas Complex But Does Not Block DNA Binding. J.Mol.Biol., 433:166759-166759, 2021 Cited by PubMed Abstract: Anti-CRISPRs are protein inhibitors of CRISPR-Cas systems. They are produced by phages and other mobile genetic elements to evade CRISPR-Cas-mediated destruction. Anti-CRISPRs are remarkably diverse in sequence, structure, and functional mechanism; thus, structural and mechanistic investigations of anti-CRISPRs continue to yield exciting new insights. In this study, we used nuclear magnetic resonance (NMR) spectroscopy to determine the solution structure of AcrIE2, an anti-CRISPR that inhibits the type I-E CRISPR-Cas system of Pseudomonas aeruginosa. Guided by the structure, we used site-directed mutagenesis to identify key residues that are required for AcrIE2 function. Using affinity purification experiments, we found that AcrIE2 binds the type I-E CRISPR-Cas complex (Cascade). In vivo transcriptional assays, in which Cascade was targeted to promoter regions, demonstrated that Cascade still binds to DNA in the presence of AcrIE2. This is the first instance of a type I anti-CRISPR that binds to a CRISPR-Cas complex but does not prevent DNA-binding. Another unusual property of AcrIE2 is that the effect of Cascade:AcrIE2 complex binding to promoter regions varied depending on the position of the binding site. Most surprisingly, Cascade:AcrIE2 binding led to transcriptional activation in some cases rather than repression, which did not occur when Cascade alone bound to the same sites. We conclude that AcrIE2 operates through a distinct mechanism compared to other type I anti-CRISPRs. While AcrIE2 does not prevent Cascade from binding DNA, it likely blocks subsequent recruitment of the Cas3 nuclease to Cascade thereby preventing DNA cleavage. PubMed: 33338493DOI: 10.1016/j.jmb.2020.166759 PDB entries with the same primary citation |
| Experimental method | SOLUTION NMR |
Structure validation
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