- EMDB-8660: Spacer capture and integration by a type I-F Cas1:Cas2-3 CRISPR a... -
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基本情報
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データベース: EMDB / ID: EMD-8660
タイトル
Spacer capture and integration by a type I-F Cas1:Cas2-3 CRISPR adaptation complex
マップデータ
Spacer capture and integration by a type I-F Cas1:Cas2-3 CRISPR adaptation complex
試料
複合体: type I-F Cas1:Cas2-3 CRISPR adaptation complex
機能・相同性
機能・相同性情報
maintenance of CRISPR repeat elements / DNA endonuclease activity / defense response to virus / 加水分解酵素; エステル加水分解酵素 / DNA binding / metal ion binding 類似検索 - 分子機能
CRISPR-associated protein Cas1, YPEST subtype / CRISPR-associated endonuclease Cas1, N-terminal domain / CRISPR-associated protein Cas1 / CRISPR-associated endonuclease Cas1, C-terminal domain / CRISPR associated protein Cas1 類似検索 - ドメイン・相同性
ジャーナル: Proc Natl Acad Sci U S A / 年: 2017 タイトル: Spacer capture and integration by a type I-F Cas1-Cas2-3 CRISPR adaptation complex. 著者: Robert D Fagerlund / Max E Wilkinson / Oleg Klykov / Arjan Barendregt / F Grant Pearce / Sebastian N Kieper / Howard W R Maxwell / Angela Capolupo / Albert J R Heck / Kurt L Krause / Mihnea ...著者: Robert D Fagerlund / Max E Wilkinson / Oleg Klykov / Arjan Barendregt / F Grant Pearce / Sebastian N Kieper / Howard W R Maxwell / Angela Capolupo / Albert J R Heck / Kurt L Krause / Mihnea Bostina / Richard A Scheltema / Raymond H J Staals / Peter C Fineran / 要旨: CRISPR-Cas adaptive immune systems capture DNA fragments from invading bacteriophages and plasmids and integrate them as spacers into bacterial CRISPR arrays. In type I-E and II-A CRISPR-Cas systems, ...CRISPR-Cas adaptive immune systems capture DNA fragments from invading bacteriophages and plasmids and integrate them as spacers into bacterial CRISPR arrays. In type I-E and II-A CRISPR-Cas systems, this adaptation process is driven by Cas1-Cas2 complexes. Type I-F systems, however, contain a unique fusion of Cas2, with the type I effector helicase and nuclease for invader destruction, Cas3. By using biochemical, structural, and biophysical methods, we present a structural model of the 400-kDa Cas1-Cas2-3 complex from with bound protospacer substrate DNA. Two Cas1 dimers assemble on a Cas2 domain dimeric core, which is flanked by two Cas3 domains forming a groove where the protospacer binds to Cas1-Cas2. We developed a sensitive in vitro assay and demonstrated that Cas1-Cas2-3 catalyzed spacer integration into CRISPR arrays. The integrase domain of Cas1 was necessary, whereas integration was independent of the helicase or nuclease activities of Cas3. Integration required at least partially duplex protospacers with free 3'-OH groups, and leader-proximal integration was stimulated by integration host factor. In a coupled capture and integration assay, Cas1-Cas2-3 processed and integrated protospacers independent of Cas3 activity. These results provide insight into the structure of protospacer-bound type I Cas1-Cas2-3 adaptation complexes and their integration mechanism.