ジャーナル: Nat Commun / 年: 2022 タイトル: Insights into the inhibition of type I-F CRISPR-Cas system by a multifunctional anti-CRISPR protein AcrIF24. 著者: Lingguang Yang / Laixing Zhang / Peipei Yin / Hao Ding / Yu Xiao / Jianwei Zeng / Wenhe Wang / Huan Zhou / Qisheng Wang / Yi Zhang / Zeliang Chen / Maojun Yang / Yue Feng / 要旨: CRISPR-Cas systems are prokaryotic adaptive immune systems and phages use anti-CRISPR proteins (Acrs) to counteract these systems. Here, we report the structures of AcrIF24 and its complex with the ...CRISPR-Cas systems are prokaryotic adaptive immune systems and phages use anti-CRISPR proteins (Acrs) to counteract these systems. Here, we report the structures of AcrIF24 and its complex with the crRNA-guided surveillance (Csy) complex. The HTH motif of AcrIF24 can bind the Acr promoter region and repress its transcription, suggesting its role as an Aca gene in self-regulation. AcrIF24 forms a homodimer and further induces dimerization of the Csy complex. Apart from blocking the hybridization of target DNA to the crRNA, AcrIF24 also induces the binding of non-sequence-specific dsDNA to the Csy complex, similar to AcrIF9, although this binding seems to play a minor role in AcrIF24 inhibitory capacity. Further structural and biochemical studies of the Csy-AcrIF24-dsDNA complexes and of AcrIF24 mutants reveal that the HTH motif of AcrIF24 and the PAM recognition loop of the Csy complex are structural elements essential for this non-specific dsDNA binding. Moreover, AcrIF24 and AcrIF9 display distinct characteristics in inducing non-specific DNA binding. Together, our findings highlight a multifunctional Acr and suggest potential wide distribution of Acr-induced non-specific DNA binding.
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B: CRISPR type I-F/YPEST-associated protein Csy2 C: CRISPR-associated protein Csy3 D: CRISPR-associated protein Csy3 E: CRISPR-associated protein Csy3 F: CRISPR-associated protein Csy3 G: CRISPR-associated protein Csy3 H: CRISPR-associated protein Csy3 I: Type I-F CRISPR-associated endoribonuclease Cas6/Csy4 L: CRISPR type I-F/YPEST-associated protein Csy2 M: CRISPR-associated protein Csy3 N: CRISPR-associated protein Csy3 O: CRISPR-associated protein Csy3 P: CRISPR-associated protein Csy3 Q: CRISPR-associated protein Csy3 R: CRISPR-associated protein Csy3 S: Type I-F CRISPR-associated endoribonuclease Cas6/Csy4 U: AcrIF24 V: AcrIF24 J: RNA (60-MER) T: RNA (60-MER) A: Type I-F CRISPR-associated protein Csy1 X: Type I-F CRISPR-associated protein Csy1 Y: DNA (5'-D(P*GP*GP*AP*TP*GP*GP*CP*TP*TP*CP*C)-3') Z: DNA-r K: DNA (5'-D(P*GP*GP*AP*TP*GP*GP*CP*TP*TP*CP*C)-3') W: DNA-r