Fig. 9: AcrIF24 and AcrIF9 show different characteristics in inducing non-specific DNA binding. | Nature Communications

Fig. 9: AcrIF24 and AcrIF9 show different characteristics in inducing non-specific DNA binding.

From: Insights into the inhibition of type I-F CRISPR-Cas system by a multifunctional anti-CRISPR protein AcrIF24

Fig. 9

a–b EMSA used to test the non-specific DNA binding affinity and mode of Csy-AcrIF24 and Csy-AcrIF9 using dsDNASP/dsDNANS (a), or ssDNASP/ssDNANS (b). Reactions were performed as in Fig. 5c. c EMSA used to test the binding ability of AcrIF24 and AcrIF9 using dsDNASP or dsDNANS. Reactions were performed as in Fig. 5c. d Structural superimposition of the Csy-AcrIF24-dsDNASP and Csy-AcrIF9-dsDNANS complexes. Only the Cas8f, Acr, and dsDNA regions are shown. K247 of Cas8f and the residues involved in non-specific DNA binding of AcrIF9 and AcrIF24 are shown in the sphere model. e EMSA was used to test the non-specific DNA binding of Csy-AcrIF9 with mutations in AcrIF9 or Csy using 3′ FAM-labeled dsDNASP or 5′ FAM-labeled dsDNANS. Reactions were performed as in Fig. 5c. Please see the left two panels of Fig. 8c for the WT control of the experiments using 3′ FAM dsDNASP 15 bp and 25 bp, respectively. Please see the left two panels of Fig. 8b for the WT control of the experiments using 5′ FAM dsDNASP 15 bp and 25 bp, respectively.

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