6DD5

Crystal Structure of the Cas6 Domain of Marinomonas mediterranea MMB-1 Cas6-RT-Cas1 Fusion Protein

6DD5 の概要

• エントリーDOI: 10.2210/pdb6dd5/pdb
• 関連するBIRD辞書のPRD_ID: PRD_900001
• 分子名称: MMB-1 Cas6 Fused to Maltose Binding Protein,CRISPR-associated endonuclease Cas1, alpha-D-glucopyranose-(1-4)-alpha-D-glucopyranose, GLYCEROL, ... (4 entities in total)
• 機能のキーワード: crispr cas protein, endoribonuclease, hydrolase
• 由来する生物種: Escherichia coli O157:H7; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 153068.14

構造登録者

Stamos, J.L.,Mohr, G.,Silas, S.,Makarova, K.S.,Markham, L.M.,Yao, J.,Lucas-Elio, P.,Sanchez-Amat, A.,Fire, A.Z.,Koonin, E.V.,Lambowitz, A.M. (登録日: 2018-05-09, 公開日: 2018-10-17, 最終更新日: 2023-10-11)

主引用文献

Mohr, G.,Silas, S.,Stamos, J.L.,Makarova, K.S.,Markham, L.M.,Yao, J.,Lucas-Elio, P.,Sanchez-Amat, A.,Fire, A.Z.,Koonin, E.V.,Lambowitz, A.M.
A Reverse Transcriptase-Cas1 Fusion Protein Contains a Cas6 Domain Required for Both CRISPR RNA Biogenesis and RNA Spacer Acquisition.
Mol. Cell, 72:700-, 2018

PubMed Abstract: Prokaryotic CRISPR-Cas systems provide adaptive immunity by integrating portions of foreign nucleic acids (spacers) into genomic CRISPR arrays. Cas6 proteins then process CRISPR array transcripts into spacer-derived RNAs (CRISPR RNAs; crRNAs) that target Cas nucleases to matching invaders. We find that a Marinomonas mediterranea fusion protein combines three enzymatic domains (Cas6, reverse transcriptase [RT], and Cas1), which function in both crRNA biogenesis and spacer acquisition from RNA and DNA. We report a crystal structure of this divergent Cas6, identify amino acids required for Cas6 activity, show that the Cas6 domain is required for RT activity and RNA spacer acquisition, and demonstrate that CRISPR-repeat binding to Cas6 regulates RT activity. Co-evolution of putative interacting surfaces suggests a specific structural interaction between the Cas6 and RT domains, and phylogenetic analysis reveals repeated, stable association of free-standing Cas6s with CRISPR RTs in multiple microbial lineages, indicating that a functional interaction between these proteins preceded evolution of the fusion.

PubMed: 30344094
DOI: 10.1016/j.molcel.2018.09.013

実験手法

X-RAY DIFFRACTION (2.85 Å)