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5XUT

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

Summary for 5XUT
Entry DOI10.2210/pdb5xut/pdb
DescriptorLbCpf1, crRNA, DNA (29-MER), ... (8 entities in total)
Functional Keywordsnuclease, hydrolase-rna-dna complex, hydrolase/rna/dna
Biological sourceLachnospiraceae bacterium ND2006
More
Total number of polymer chains4
Total formula weight168777.19
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 citationYamano, 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
Cited by
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
PDB entries with the same primary citation
Experimental method
X-RAY DIFFRACTION (2.4 Å)
Structure validation

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