5XUU

Crystal structure of Lachnospiraceae bacterium ND2006 Cpf1 in complex with crRNA and target DNA (TCCA PAM)

Summary for 5XUU

• Entry DOI: 10.2210/pdb5xuu/pdb
• Descriptor: LbCpf1, crRNA, DNA (29-MER), ... (8 entities in total)
• Functional Keywords: nuclease, hydrolase-rna-dna complex, hydrolase/rna/dna
• Biological source: Lachnospiraceae bacterium ND2006; More
• Total number of polymer chains: 4
• Total formula weight: 168778.18

Authors

Yamano, T.,Nishimasu, H.,Ishitani, R.,Nureki, O. (deposition date: 2017-06-26, release date: 2017-08-09, Last modification date: 2023-11-22)

Primary citation

Yamano, T.,Zetsche, B.,Ishitani, R.,Zhang, F.,Nishimasu, H.,Nureki, O.
Structural Basis for the Canonical and Non-canonical PAM Recognition by CRISPR-Cpf1.
Mol. Cell, 67:633-645.e3, 2017

PubMed Abstract: The RNA-guided Cpf1 (also known as Cas12a) nuclease associates with a CRISPR RNA (crRNA) and cleaves the double-stranded DNA target complementary to the crRNA guide. The two Cpf1 orthologs from Acidaminococcus sp. (AsCpf1) and Lachnospiraceae bacterium (LbCpf1) have been harnessed for eukaryotic genome editing. Cpf1 requires a specific nucleotide sequence, called a protospacer adjacent motif (PAM), for target recognition. Besides the canonical TTTV PAM, Cpf1 recognizes suboptimal C-containing PAMs. Here, we report four crystal structures of LbCpf1 in complex with the crRNA and its target DNA containing either TTTA, TCTA, TCCA, or CCCA as the PAM. These structures revealed that, depending on the PAM sequences, LbCpf1 undergoes conformational changes to form altered interactions with the PAM-containing DNA duplexes, thereby achieving the relaxed PAM recognition. Collectively, the present structures advance our mechanistic understanding of the PAM-dependent, crRNA-guided DNA cleavage by the Cpf1 family nucleases.

PubMed: 28781234
DOI: 10.1016/j.molcel.2017.06.035

Experimental method

X-RAY DIFFRACTION (2.5 Å)