PRJNA518078
Cas4-Cas1 fusions drive efficient PAM selection and control CRISPR adaptation
Microbes have the unique ability to acquire immunological memories from mobile genetic invaders to protect themselves from predation. To confer CRISPR resistance, new spacers need to be compatible with a targeting requirement in the invader's DNA called the protospacer adjacent motif (PAM). Many CRISPR systems encode Cas4 proteins to ensure new spacers are integrated that meet this targeting prerequisite. Here we report that a gene fusion between cas4 and cas1 from the Geobacter sulfurreducens I-U CRISPR-Cas system is capable of introducing functional spacers carrying interference . Mutations of Cas4-domain catalytic residues resulted in dramatically decreasedspacer acquisition, and a loss of PAM selectivity showing that the Cas4 domain controls Cas1 activity.