5XUS

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

5XUS の概要

• エントリーDOI: 10.2210/pdb5xus/pdb
• 分子名称: LbCpf1, crRNA, DNA (29-MER), ... (8 entities in total)
• 機能のキーワード: nuclease, hydrolase-rna-dna complex, hydrolase/rna/dna
• 由来する生物種: Lachnospiraceae bacterium ND2006; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 168846.48

構造登録者

Yamano, T.,Nishimasu, H.,Ishitani, R.,Nureki, O. (登録日: 2017-06-26, 公開日: 2017-08-09, 最終更新日: 2024-11-06)

主引用文献

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

実験手法

X-RAY DIFFRACTION (2.5 Å)