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TitleAcrIF5 specifically targets DNA-bound CRISPR-Cas surveillance complex for inhibition.
Journal, issue, pagesNat Chem Biol, Vol. 18, Issue 6, Page 670-677, Year 2022
Publish dateMar 17, 2022
AuthorsYongchao Xie / Laixing Zhang / Zhengyu Gao / Peipei Yin / Hao Wang / Hang Li / Zeliang Chen / Yi Zhang / Maojun Yang / Yue Feng /
PubMed AbstractCRISPR-Cas systems are prokaryotic antiviral systems, and phages use anti-CRISPR proteins (Acrs) to inactivate these systems. Here we present structural and functional analyses of AcrIF5, exploring ...CRISPR-Cas systems are prokaryotic antiviral systems, and phages use anti-CRISPR proteins (Acrs) to inactivate these systems. Here we present structural and functional analyses of AcrIF5, exploring its unique anti-CRISPR mechanism. AcrIF5 shows binding specificity only for the target DNA-bound form of the crRNA-guided surveillance (Csy) complex, but not the apo Csy complex from the type I-F CRISPR-Cas system. We solved the structure of the Csy-dsDNA-AcrIF5 complex, revealing that the conformational changes of the Csy complex caused by dsDNA binding dictate the binding specificity for the Csy-dsDNA complex by AcrIF5. Mechanistically, five AcrIF5 molecules bind one Csy-dsDNA complex, which destabilizes the helical bundle domain of Cas8f, thus preventing subsequent Cas2/3 recruitment. AcrIF5 exists in symbiosis with AcrIF3, which blocks Cas2/3 recruitment. This attack on the recruitment event stands in contrast to the conventional mechanisms of blocking binding of target DNA. Overall, our study reveals an unprecedented mechanism of CRISPR-Cas inhibition by AcrIF5.
External linksNat Chem Biol / PubMed:35301482
MethodsEM (single particle) / X-ray diffraction
Resolution3.2 - 3.52 Å
Structure data

31265

7eqg

EMDB-31265, PDB-7eqg:
Structure of Csy-AcrIF5
Method: EM (single particle) / Resolution: 3.2 Å

PDB-7f45:
Structure of an Anti-CRISPR protein
Method: X-RAY DIFFRACTION / Resolution: 3.52 Å

Source
  • pseudomonas aeruginosa (bacteria)
KeywordsIMMUNE SYSTEM/RNA/DNA / complex / inhibitor / IMMUNE SYSTEM / IMMUNE SYSTEM-RNA-DNA complex / monomer / mixed alpha-beta structure

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