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TitleInsights into the dual functions of AcrIF14 during the inhibition of type I-F CRISPR-Cas surveillance complex.
Journal, issue, pagesNucleic Acids Res, Vol. 49, Issue 17, Page 10178-10191, Year 2021
Publish dateSep 27, 2021
AuthorsXi Liu / Laixing Zhang / Yu Xiu / Teng Gao / Ling Huang / Yongchao Xie / Lingguang Yang / Wenhe Wang / Peiyi Wang / Yi Zhang / Maojun Yang / Yue Feng /
PubMed AbstractCRISPR-Cas systems are bacterial adaptive immune systems, and phages counteract these systems using many approaches such as producing anti-CRISPR (Acr) proteins. Here, we report the structures of ...CRISPR-Cas systems are bacterial adaptive immune systems, and phages counteract these systems using many approaches such as producing anti-CRISPR (Acr) proteins. Here, we report the structures of both AcrIF14 and its complex with the crRNA-guided surveillance (Csy) complex. Our study demonstrates that apart from interacting with the Csy complex to block the hybridization of target DNA to the crRNA, AcrIF14 also endows the Csy complex with the ability to interact with non-sequence-specific dsDNA as AcrIF9 does. Further structural studies of the Csy-AcrIF14-dsDNA complex and biochemical studies uncover that the PAM recognition loop of the Cas8f subunit of the Csy complex and electropositive patches within the N-terminal domain of AcrIF14 are essential for the non-sequence-specific dsDNA binding to the Csy-AcrIF14 complex, which is different from the mechanism of AcrIF9. Our findings highlight the prevalence of Acr-induced non-specific DNA binding and shed light on future studies into the mechanisms of such Acr proteins.
External linksNucleic Acids Res / PubMed:34432044 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution1.96 - 3.43 Å
Structure data

EMDB-31058, PDB-7ecv:
The Csy-AcrIF14 complex
Method: EM (single particle) / Resolution: 3.43 Å

EMDB-31059, PDB-7ecw:
The Csy-AcrIF14-dsDNA complex
Method: EM (single particle) / Resolution: 3.1 Å

PDB-7du0:
Structure of an type I-F anti-crispr protein
Method: X-RAY DIFFRACTION / Resolution: 1.96 Å

Chemicals

ChemComp-HOH:
WATER

Source
  • pseudomonas aeruginosa (bacteria)
  • moraxella phage mcat5 (virus)
KeywordsVIRAL PROTEIN / monomer / two-domain / IMMUNE SYSTEM/RNA / inhibitor / complex / IMMUNE SYSTEM / IMMUNE SYSTEM-RNA complex / IMMUNE SYSTEM/RNA/DNA / IMMUNE SYSTEM-RNA-DNA complex

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