+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-25421 | |||||||||||||||
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タイトル | Pre translocation intermediate stalled with viomycin and bound with EF-G in a GDP and Pi state (Structure III-vio) | |||||||||||||||
マップデータ | beamtilt corrected map used in refinement of III-vio | |||||||||||||||
試料 |
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機能・相同性 | 機能・相同性情報 negative regulation of cytoplasmic translational initiation / stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity ...negative regulation of cytoplasmic translational initiation / stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / positive regulation of ribosome biogenesis / negative regulation of cytoplasmic translation / translation elongation factor activity / translational termination / four-way junction DNA binding / DnaA-L2 complex / translation repressor activity / negative regulation of DNA-templated DNA replication initiation / negative regulation of translational initiation / regulation of mRNA stability / mRNA regulatory element binding translation repressor activity / ribosome assembly / assembly of large subunit precursor of preribosome / positive regulation of RNA splicing / transcription elongation factor complex / cytosolic ribosome assembly / regulation of DNA-templated transcription elongation / DNA endonuclease activity / response to reactive oxygen species / transcription antitermination / regulation of cell growth / translational initiation / DNA-templated transcription termination / maintenance of translational fidelity / response to radiation / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / ribosome biogenesis / ribosome binding / regulation of translation / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / transferase activity / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / tRNA binding / molecular adaptor activity / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / response to antibiotic / negative regulation of DNA-templated transcription / GTPase activity / mRNA binding / GTP binding / DNA binding / RNA binding / zinc ion binding / membrane / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||||||||
生物種 | Escherichia coli K-12 (大腸菌) / Streptomyces californicus (バクテリア) | |||||||||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.2 Å | |||||||||||||||
データ登録者 | Carbone CE / Korostelev AA | |||||||||||||||
資金援助 | チェコ, 4件
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引用 | ジャーナル: Nat Commun / 年: 2021 タイトル: Time-resolved cryo-EM visualizes ribosomal translocation with EF-G and GTP. 著者: Christine E Carbone / Anna B Loveland / Howard B Gamper / Ya-Ming Hou / Gabriel Demo / Andrei A Korostelev / 要旨: During translation, a conserved GTPase elongation factor-EF-G in bacteria or eEF2 in eukaryotes-translocates tRNA and mRNA through the ribosome. EF-G has been proposed to act as a flexible motor that ...During translation, a conserved GTPase elongation factor-EF-G in bacteria or eEF2 in eukaryotes-translocates tRNA and mRNA through the ribosome. EF-G has been proposed to act as a flexible motor that propels tRNA and mRNA movement, as a rigid pawl that biases unidirectional translocation resulting from ribosome rearrangements, or by various combinations of motor- and pawl-like mechanisms. Using time-resolved cryo-EM, we visualized GTP-catalyzed translocation without inhibitors, capturing elusive structures of ribosome•EF-G intermediates at near-atomic resolution. Prior to translocation, EF-G binds near peptidyl-tRNA, while the rotated 30S subunit stabilizes the EF-G GTPase center. Reverse 30S rotation releases Pi and translocates peptidyl-tRNA and EF-G by ~20 Å. An additional 4-Å translocation initiates EF-G dissociation from a transient ribosome state with highly swiveled 30S head. The structures visualize how nearly rigid EF-G rectifies inherent and spontaneous ribosomal dynamics into tRNA-mRNA translocation, whereas GTP hydrolysis and Pi release drive EF-G dissociation. | |||||||||||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | EMマップ: SurfViewMolmilJmol/JSmol |
添付画像 |
-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_25421.map.gz | 317.9 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-25421-v30.xml emd-25421.xml | 75.1 KB 75.1 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_25421.png | 104.6 KB | ||
その他 | emd_25421_additional_1.map.gz emd_25421_half_map_1.map.gz emd_25421_half_map_2.map.gz | 320.6 MB 144.1 MB 144.1 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-25421 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-25421 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_25421_validation.pdf.gz | 875.4 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_25421_full_validation.pdf.gz | 875 KB | 表示 | |
XML形式データ | emd_25421_validation.xml.gz | 17.6 KB | 表示 | |
CIF形式データ | emd_25421_validation.cif.gz | 20.8 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25421 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25421 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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「今月の分子」の関連する項目 |
-マップ
ファイル | ダウンロード / ファイル: emd_25421.map.gz / 形式: CCP4 / 大きさ: 343 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | beamtilt corrected map used in refinement of III-vio | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 0.87 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
-追加マップ: beamtilt corrected map of III-vio that was blocfiltered...
ファイル | emd_25421_additional_1.map | ||||||||||||
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注釈 | beamtilt corrected map of III-vio that was blocfiltered and sharpened to -50 at 3.2A | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: half map 1
ファイル | emd_25421_half_map_1.map | ||||||||||||
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注釈 | half map 1 | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: half map 2
ファイル | emd_25421_half_map_2.map | ||||||||||||
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注釈 | half map 2 | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-試料の構成要素
+全体 : Pre translocation intermediate stalled with viomycin and bound wi...
+超分子 #1: Pre translocation intermediate stalled with viomycin and bound wi...
+分子 #1: 23S rRNA
+分子 #2: 5S rRNA
+分子 #41: 16S rRNA
+分子 #42: mRNA
+分子 #57: tRNA fMet
+分子 #58: tRNA Pro
+分子 #3: 50S ribosomal protein L2
+分子 #4: 50S ribosomal protein L3
+分子 #5: 50S ribosomal protein L4
+分子 #6: 50S ribosomal protein L5
+分子 #7: 50S ribosomal protein L6
+分子 #8: 50S ribosomal protein L9
+分子 #9: 50S ribosomal protein L1
+分子 #10: 50S ribosomal protein L11
+分子 #11: 50S ribosomal protein L13
+分子 #12: 50S ribosomal protein L14
+分子 #13: 50S ribosomal protein L15
+分子 #14: 50S ribosomal protein L16
+分子 #15: 50S ribosomal protein L17
+分子 #16: 50S ribosomal protein L18
+分子 #17: 50S ribosomal protein L19
+分子 #18: 50S ribosomal protein L20
+分子 #19: 50S ribosomal protein L21
+分子 #20: 50S ribosomal protein L22
+分子 #21: 50S ribosomal protein L23
+分子 #22: 50S ribosomal protein L24
+分子 #23: 50S ribosomal protein L25
+分子 #24: 50S ribosomal protein L27
+分子 #25: 50S ribosomal protein L28
+分子 #26: 50S ribosomal protein L29
+分子 #27: 50S ribosomal protein L30
+分子 #28: 50S ribosomal protein L31
+分子 #29: 50S ribosomal protein L32
+分子 #30: 50S ribosomal protein L33
+分子 #31: 50S ribosomal protein L34
+分子 #32: 50S ribosomal protein L35
+分子 #33: 50S ribosomal protein L36
+分子 #34: 30S ribosomal protein S3
+分子 #35: 30S ribosomal protein S7
+分子 #36: 30S ribosomal protein S9
+分子 #37: 30S ribosomal protein S10
+分子 #38: 30S ribosomal protein S13
+分子 #39: 30S ribosomal protein S14
+分子 #40: 30S ribosomal protein S19
+分子 #43: 30S ribosomal protein S2
+分子 #44: 30S ribosomal protein S4
+分子 #45: 30S ribosomal protein S5
+分子 #46: 30S ribosomal protein S6
+分子 #47: 30S ribosomal protein S8
+分子 #48: 30S ribosomal protein S11
+分子 #49: 30S ribosomal protein S12
+分子 #50: 30S ribosomal protein S15
+分子 #51: 30S ribosomal protein S16
+分子 #52: 30S ribosomal protein S17
+分子 #53: 30S ribosomal protein S18
+分子 #54: 30S ribosomal protein S20
+分子 #55: 30S ribosomal protein S21
+分子 #56: Elongation factor G
+分子 #59: Viomycin
+分子 #60: GUANOSINE-5'-DIPHOSPHATE
+分子 #61: PHOSPHATE ION
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
緩衝液 | pH: 7.5 |
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凍結 | 凍結剤: ETHANE |
-電子顕微鏡法
顕微鏡 | FEI TALOS ARCTICA |
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撮影 | フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 平均電子線量: 30.5 e/Å2 |
電子線 | 加速電圧: 200 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 1.5 µm / 最小 デフォーカス(公称値): 0.5 µm |
実験機器 | モデル: Talos Arctica / 画像提供: FEI Company |