Research Publications

Two Distinct DNA Binding Modes Guide Dual Roles of a CRISPR-Cas Protein Complex

Year of publication
2015
Author
chirlmin
Date
01 Apr 2015 19:28

Molecular Cell. 2015 Mar 4.

doi: 10.1016/j.molcel.2015.01.028

Two Distinct DNA Binding Modes Guide Dual Roles of a CRISPR-Cas Protein Complex.
Blosser TR, Loeff L, Westra ER, Vlot M, Künne T, Sobota M, Dekker C, Brouns SJ, Joo C.

Small RNA-guided protein complexes play an essential role in CRISPR-mediated immunity in prokaryotes. While these complexes initiate interference by flagging cognate invader DNA for destruction, recent evidence has implicated their involvement in new CRISPR memory formation, called priming, against mutated invader sequences. The mechanism by which the target recognition complex mediates these disparate responses-interference and priming-remains poorly understood. Using single-molecule FRET, we visualize how bona fide and mutated targets are differentially probed by E. coli Cascade. We observe that the recognition of bona fide targets is an ordered process that is tightly controlled for high fidelity. Mutated targets are recognized with low fidelity, which is featured by short-lived and PAM- and seed-independent binding by any segment of the crRNA. These dual roles of Cascade in immunity with distinct fidelities underpin CRISPR-Cas robustness, allowing for efficient degradation of bona fide targets and priming of mutated DNA targets.

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