7KFU

Cas6-RT-Cas1--Cas2 complex

7KFU の概要

• エントリーDOI: 10.2210/pdb7kfu/pdb
• 関連するPDBエントリー: 7KFT
• EMDBエントリー: 22855 22856
• 分子名称: Cas2, Cas6-RT-Cas1 (2 entities in total)
• 機能のキーワード: complex, hydrolase, crispr cas protein, reverse transcriptase
• 由来する生物種: Thiomicrospira sp.; 詳細
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 474644.18

構造登録者

Hoel, C.M.,Wang, J.Y.,Doudna, J.A.,Brohawn, S.G. (登録日: 2020-10-14, 公開日: 2021-03-31, 最終更新日: 2024-03-06)

主引用文献

Wang, J.Y.,Hoel, C.M.,Al-Shayeb, B.,Banfield, J.F.,Brohawn, S.G.,Doudna, J.A.
Structural coordination between active sites of a CRISPR reverse transcriptase-integrase complex.
Nat Commun, 12:2571-2571, 2021

PubMed Abstract: CRISPR-Cas systems provide adaptive immunity in bacteria and archaea, beginning with integration of foreign sequences into the host CRISPR genomic locus and followed by transcription and maturation of CRISPR RNAs (crRNAs). In some CRISPR systems, a reverse transcriptase (RT) fusion to the Cas1 integrase and Cas6 maturase creates a single protein that enables concerted sequence integration and crRNA production. To elucidate how the RT-integrase organizes distinct enzymatic activities, we present the cryo-EM structure of a Cas6-RT-Cas1-Cas2 CRISPR integrase complex. The structure reveals a heterohexamer in which the RT directly contacts the integrase and maturase domains, suggesting functional coordination between all three active sites. Together with biochemical experiments, our data support a model of sequential enzymatic activities that enable CRISPR sequence acquisition from RNA and DNA substrates. These findings highlight an expanded capacity of some CRISPR systems to acquire diverse sequences that direct CRISPR-mediated interference.

PubMed: 33958590
DOI: 10.1038/s41467-021-22900-y

実験手法

ELECTRON MICROSCOPY (3.9 Å)