5U30

Crystal structure of AacC2c1-sgRNA-extended target DNA ternary complex

Summary for 5U30

• Entry DOI: 10.2210/pdb5u30/pdb
• Related: 5U31 5U33 5U34
• Descriptor: CRISPR-associated endonuclease C2c1, sgRNA, Target DNA strand, ... (6 entities in total)
• Functional Keywords: type v crispr-cas endonculease: c2c1: structure: binary complex with sgrna: ternary complex with added dna: ruvc catalytic pocket: sequence-specific pam recognition: genome editing tool, hydrolase-dna complex, hydrolase/dna
• Biological source: Alicyclobacillus acidoterrestris; More
• Total number of polymer chains: 4
• Total formula weight: 182271.81

Authors

Yang, H.,Gao, P.,Rajashankar, K.R.,Patel, D.J. (deposition date: 2016-12-01, release date: 2017-01-25, Last modification date: 2024-12-25)

Primary citation

Yang, H.,Gao, P.,Rajashankar, K.R.,Patel, D.J.
PAM-Dependent Target DNA Recognition and Cleavage by C2c1 CRISPR-Cas Endonuclease.
Cell, 167:1814-1828.e12, 2016

PubMed Abstract: C2c1 is a newly identified guide RNA-mediated type V-B CRISPR-Cas endonuclease that site-specifically targets and cleaves both strands of target DNA. We have determined crystal structures of Alicyclobacillus acidoterrestris C2c1 (AacC2c1) bound to sgRNA as a binary complex and to target DNAs as ternary complexes, thereby capturing catalytically competent conformations of AacC2c1 with both target and non-target DNA strands independently positioned within a single RuvC catalytic pocket. Moreover, C2c1-mediated cleavage results in a staggered seven-nucleotide break of target DNA. crRNA adopts a pre-ordered five-nucleotide A-form seed sequence in the binary complex, with release of an inserted tryptophan, facilitating zippering up of 20-bp guide RNA:target DNA heteroduplex on ternary complex formation. Notably, the PAM-interacting cleft adopts a "locked" conformation on ternary complex formation. Structural comparison of C2c1 ternary complexes with their Cas9 and Cpf1 counterparts highlights the diverse mechanisms adopted by these distinct CRISPR-Cas systems, thereby broadening and enhancing their applicability as genome editing tools.

PubMed: 27984729
DOI: 10.1016/j.cell.2016.11.053

Experimental method

X-RAY DIFFRACTION (2.92 Å)