5B43

Crystal structure of Acidaminococcus sp. Cpf1 in complex with crRNA and target DNA

Summary for 5B43

• Entry DOI: 10.2210/pdb5b43/pdb
• Descriptor: CRISPR-associated endonuclease Cpf1, RNA (43-MER), DNA (34-MER), ... (7 entities in total)
• Functional Keywords: nuclease, hydrolase-rna-dna complex, hydrolase/rna/dna
• Biological source: Acidaminococcus sp. BV3L6; More
• Total number of polymer chains: 4
• Total formula weight: 179105.26

Authors

Yamano, T.,Nishimasu, H.,Hirano, H.,Nakane, T.,Ishitani, R.,Nureki, O. (deposition date: 2016-03-30, release date: 2016-05-04, Last modification date: 2024-03-20)

Primary citation

Yamano, T.,Nishimasu, H.,Zetsche, B.,Hirano, H.,Slaymaker, I.M.,Li, Y.,Fedorova, I.,Nakane, T.,Makarova, K.S.,Koonin, E.V.,Ishitani, R.,Zhang, F.,Nureki, O.
Crystal Structure of Cpf1 in Complex with Guide RNA and Target DNA
Cell, 165:949-962, 2016

PubMed Abstract: Cpf1 is an RNA-guided endonuclease of a type V CRISPR-Cas system that has been recently harnessed for genome editing. Here, we report the crystal structure of Acidaminococcus sp. Cpf1 (AsCpf1) in complex with the guide RNA and its target DNA at 2.8 Å resolution. AsCpf1 adopts a bilobed architecture, with the RNA-DNA heteroduplex bound inside the central channel. The structural comparison of AsCpf1 with Cas9, a type II CRISPR-Cas nuclease, reveals both striking similarity and major differences, thereby explaining their distinct functionalities. AsCpf1 contains the RuvC domain and a putative novel nuclease domain, which are responsible for cleaving the non-target and target strands, respectively, and for jointly generating staggered DNA double-strand breaks. AsCpf1 recognizes the 5'-TTTN-3' protospacer adjacent motif by base and shape readout mechanisms. Our findings provide mechanistic insights into RNA-guided DNA cleavage by Cpf1 and establish a framework for rational engineering of the CRISPR-Cpf1 toolbox.

PubMed: 27114038
DOI: 10.1016/j.cell.2016.04.003

Experimental method

X-RAY DIFFRACTION (2.8 Å)