Structure Studies of the CRISPR-Csm Complex Reveal Mechanism of Co-transcriptional Interference

2018
Summary Csm, a type III-A CRISPR-Cas interference complex, is a CRISPR RNA(crRNA)-guided RNase that also possesses target RNA-dependent DNase and cyclic oligoadenylate (cOA) synthetase activities. However, the structural features allowing target RNA-binding-dependent activation of DNA cleavage and cOA generation remain unknown. Here, we report the structure of Csm in complex with crRNA together with structures of cognateor non- cognatetarget RNAbound Csm complexes. We show that depending on complementarity with the 5′ tag of crRNA, the 3′ anti-tag region of target RNAbinds at two distinct sites of the Csm complex. Importantly, the interaction between the non-complementary anti-tag region of cognatetarget RNAand Csm1 induces a conformational change at the Csm1 subunit that allosterically activates DNA cleavage and cOA generation. Together, our structural studies provide crucial insights into the mechanistic processes required for crRNA-meditated sequence-specific RNAcleavage, RNAtarget-dependent non-specific DNA cleavage, and cOA generation.
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