- EMDB-2418: Structure and activity of an RNA-targeting Type III-B CRISPER-Cas... -
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IDまたはキーワード:
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基本情報
登録情報
データベース: EMDB / ID: EMD-2418
タイトル
Structure and activity of an RNA-targeting Type III-B CRISPER-Cas complex
マップデータ
Reconstruction of Type III-B CRISPR-Cas complex from Thermus thermophilus
試料
試料: Type III-B CRISPR-Cas complex of Thermus thermophilus
タンパク質・ペプチド: CRISPR-associated protein TM1791
タンパク質・ペプチド: CRISPR system Cmr subunit Cmr5
タンパク質・ペプチド: CRISPR-associated protein Cmr3
タンパク質・ペプチド: CRISPR-associated protein Crm2
タンパク質・ペプチド: CRISPR-associated RAMP Cmr4
タンパク質・ペプチド: CRISPR-associated protein TM1795
RNA: CRISPR RNA
キーワード
CRISPR-Cas / RNA-targeting / Cmr complex
機能・相同性
機能・相同性情報
regulation of defense response to virus / defense response to virus / cytoplasm 類似検索 - 分子機能
CRISPR-associated protein (Cas_Cmr5) / CRISPR-associated protein, TM1791 / CRISPR-associated protein, TM1791 / CRISPR-associated protein TM1795 / CRISPR-associated protein TM1795 / CRISPR-associated protein, Cmr3 / CRISPR-associated protein, Cmr3 / CRISPR-associated protein (Cas_Cmr3) / CRISPR-associated protein, Cmr5 / CRISPR-associated RAMP Cmr4 ...CRISPR-associated protein (Cas_Cmr5) / CRISPR-associated protein, TM1791 / CRISPR-associated protein, TM1791 / CRISPR-associated protein TM1795 / CRISPR-associated protein TM1795 / CRISPR-associated protein, Cmr3 / CRISPR-associated protein, Cmr3 / CRISPR-associated protein (Cas_Cmr3) / CRISPR-associated protein, Cmr5 / CRISPR-associated RAMP Cmr4 / CRISPR-associated protein, Cmr5 / CRISPR-associated RAMP Cmr4 / AF1862-like domain superfamily / CRISPR-associated protein Cmr2 / CRISPR-associated protein Cmr2 / CRISPR-associated protein Cmr2, N-terminal / CRISPR-Cas system, Cmr2 subunit, D1 domain, cysteine cluster / CRISPR-associated protein Cmr2, N-terminal / CRISPR type III-associated protein / RAMP superfamily / GGDEF domain profile. / GGDEF domain / Reverse transcriptase/Diguanylate cyclase domain 類似検索 - ドメイン・相同性
Type III-B CRISPR module RAMP protein Cmr6 / CRISPR system Cmr subunit Cmr5 / Type III-B CRISPR module RAMP protein Cmr4 / Type III-B CRISPR module RAMP protein Cmr1 / CRISPR-associated protein Cmr3 / GGDEF domain-containing protein 類似検索 - 構成要素
ジャーナル: Mol Cell / 年: 2013 タイトル: Structure and activity of the RNA-targeting Type III-B CRISPR-Cas complex of Thermus thermophilus. 著者: Raymond H J Staals / Yoshihiro Agari / Saori Maki-Yonekura / Yifan Zhu / David W Taylor / Esther van Duijn / Arjan Barendregt / Marnix Vlot / Jasper J Koehorst / Keiko Sakamoto / Akiko Masuda ...著者: Raymond H J Staals / Yoshihiro Agari / Saori Maki-Yonekura / Yifan Zhu / David W Taylor / Esther van Duijn / Arjan Barendregt / Marnix Vlot / Jasper J Koehorst / Keiko Sakamoto / Akiko Masuda / Naoshi Dohmae / Peter J Schaap / Jennifer A Doudna / Albert J R Heck / Koji Yonekura / John van der Oost / Akeo Shinkai / 要旨: The CRISPR-Cas system is a prokaryotic host defense system against genetic elements. The Type III-B CRISPR-Cas system of the bacterium Thermus thermophilus, the TtCmr complex, is composed of six ...The CRISPR-Cas system is a prokaryotic host defense system against genetic elements. The Type III-B CRISPR-Cas system of the bacterium Thermus thermophilus, the TtCmr complex, is composed of six different protein subunits (Cmr1-6) and one crRNA with a stoichiometry of Cmr112131445361:crRNA1. The TtCmr complex copurifies with crRNA species of 40 and 46 nt, originating from a distinct subset of CRISPR loci and spacers. The TtCmr complex cleaves the target RNA at multiple sites with 6 nt intervals via a 5' ruler mechanism. Electron microscopy revealed that the structure of TtCmr resembles a "sea worm" and is composed of a Cmr2-3 heterodimer "tail," a helical backbone of Cmr4 subunits capped by Cmr5 subunits, and a curled "head" containing Cmr1 and Cmr6. Despite having a backbone of only four Cmr4 subunits and being both longer and narrower, the overall architecture of TtCmr resembles that of Type I Cascade complexes.
UniProtKB: Type III-B CRISPR module RAMP protein Cmr4 / InterPro: CRISPR-associated RAMP Cmr4
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分子 #7: CRISPR-associated protein TM1795
分子
名称: CRISPR-associated protein TM1795 / タイプ: protein_or_peptide / ID: 7 / Name.synonym: Cmr1, TTHB162 / コピー数: 1 / 組換発現: No
由来(天然)
生物種: Thermus thermophilus (バクテリア)
分子量
実験値: 44 KDa
配列
UniProtKB: Type III-B CRISPR module RAMP protein Cmr1 / InterPro: CRISPR-associated protein TM1795
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分子 #2: CRISPR RNA
分子
名称: CRISPR RNA / タイプ: rna / ID: 2 / Name.synonym: crRNA / 詳細: 40-46 nucleotide endogenous crRNA bound to complex / 分類: OTHER / Structure: SINGLE STRANDED / Synthetic?: No
由来(天然)
生物種: Thermus thermophilus (バクテリア)
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実験情報
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構造解析
手法
ネガティブ染色法
解析
単粒子再構成法
試料の集合状態
particle
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試料調製
濃度
0.03 mg/mL
緩衝液
pH: 7.4 詳細: 25 mM Hepes, pH 7.5, 100 mM KCl, 1 mM TCEP, 5% glycerol
染色
タイプ: NEGATIVE 詳細: Sample applied to glow-discharged continuous carbon grids. After 1 min, grids were stained consecutively in 6 droplets of 2% (w/v) uranyl acetate solution and excess stain removed by gentle ...詳細: Sample applied to glow-discharged continuous carbon grids. After 1 min, grids were stained consecutively in 6 droplets of 2% (w/v) uranyl acetate solution and excess stain removed by gentle blotting with filter paper.
グリッド
詳細: 200 mesh Cu grid
凍結
凍結剤: NONE / 装置: OTHER
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電子顕微鏡法
顕微鏡
FEI TECNAI 20
アライメント法
Legacy - 非点収差: Objective astigmatism was corrected at 210,000 times magnification