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TitleStructural basis of AcrIF24 as an anti-CRISPR protein and transcriptional suppressor.
Journal, issue, pagesNat Chem Biol, Vol. 18, Issue 12, Page 1417-1424, Year 2022
Publish dateSep 26, 2022
AuthorsIndranil Arun Mukherjee / Clinton Gabel / Nicholas Noinaj / Joseph Bondy-Denomy / Leifu Chang /
PubMed AbstractAnti-CRISPR (Acr) proteins are encoded by phages to inactivate CRISPR-Cas systems of bacteria and archaea and are used to enhance the CRISPR toolbox for genome editing. Here we report the structure ...Anti-CRISPR (Acr) proteins are encoded by phages to inactivate CRISPR-Cas systems of bacteria and archaea and are used to enhance the CRISPR toolbox for genome editing. Here we report the structure and mechanism of AcrIF24, an Acr protein that inhibits the type I-F CRISPR-Cas system from Pseudomonas aeruginosa. AcrIF24 is a homodimer that associates with two copies of the surveillance complex (Csy) and prevents the hybridization between CRISPR RNA and target DNA. Furthermore, AcrIF24 functions as an anti-CRISPR-associated (Aca) protein to repress the transcription of the acrIF23-acrIF24 operon. Alone or in complex with Csy, AcrIF24 is capable of binding to the acrIF23-acrIF24 promoter DNA with nanomolar affinity. The structure of a Csy-AcrIF24-promoter DNA complex at 2.7 Å reveals the mechanism for transcriptional suppression. Our results reveal that AcrIF24 functions as an Acr-Aca fusion protein, and they extend understanding of the diverse mechanisms used by Acr proteins.
External linksNat Chem Biol / PubMed:36163386 / PubMed Central
MethodsEM (single particle)
Resolution2.7 - 3.6 Å
Structure data

25660

7t3j

EMDB-25660, PDB-7t3j:
Cryo-EM structure of Csy-AcrIF24
Method: EM (single particle) / Resolution: 3.2 Å

25661

7t3k

EMDB-25661, PDB-7t3k:
Cryo-EM structure of Csy-AcrIF24 dimer
Method: EM (single particle) / Resolution: 3.5 Å

25662

7t3l

EMDB-25662, PDB-7t3l:
Cryo-EM structure of Csy-AcrIF24-DNA dimer
Method: EM (single particle) / Resolution: 3.6 Å

25788

7taw

EMDB-25788, PDB-7taw:
Cryo-EM structure of the Csy-AcrIF24-promoter DNA dimer
Method: EM (single particle) / Resolution: 2.7 Å

25789

7tax

EMDB-25789, PDB-7tax:
Cryo-EM structure of the Csy-AcrIF24-promoter DNA complex
Method: EM (single particle) / Resolution: 2.8 Å

Source
  • pseudomonas (bacteria)
  • escherichia (bacteria)
KeywordsHYDROLASE/RNA / Csy / Cascade / CRISPR / anti-CRISPR / AcrIF24 / HYDROLASE-RNA complex / HYDROLASE/RNA/DNA / HYDROLASE-RNA-DNA complex

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