+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-16634 | |||||||||
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タイトル | Translocation intermediate 3 (TI-3) of 80S S. cerevisiae ribosome with ligands and eEF2 in the presence of sordarin | |||||||||
マップデータ | ||||||||||
試料 |
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キーワード | Eukaryote / ribosome / translocation / elongation | |||||||||
機能・相同性 | 機能・相同性情報 ribosomal subunit / negative regulation of glucose mediated signaling pathway / negative regulation of translational frameshifting / Negative regulators of DDX58/IFIH1 signaling / RMTs methylate histone arginines / positive regulation of translational fidelity / Protein methylation / mTORC1-mediated signalling / Protein hydroxylation / ribosome-associated ubiquitin-dependent protein catabolic process ...ribosomal subunit / negative regulation of glucose mediated signaling pathway / negative regulation of translational frameshifting / Negative regulators of DDX58/IFIH1 signaling / RMTs methylate histone arginines / positive regulation of translational fidelity / Protein methylation / mTORC1-mediated signalling / Protein hydroxylation / ribosome-associated ubiquitin-dependent protein catabolic process / GDP-dissociation inhibitor activity / nonfunctional rRNA decay / hexon binding / positive regulation of nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / pre-mRNA 5'-splice site binding / Formation of the ternary complex, and subsequently, the 43S complex / Translation initiation complex formation / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / Ribosomal scanning and start codon recognition / response to cycloheximide / Major pathway of rRNA processing in the nucleolus and cytosol / mRNA destabilization / SRP-dependent cotranslational protein targeting to membrane / GTP hydrolysis and joining of the 60S ribosomal subunit / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / Formation of a pool of free 40S subunits / negative regulation of mRNA splicing, via spliceosome / preribosome, large subunit precursor / regulation of amino acid metabolic process / L13a-mediated translational silencing of Ceruloplasmin expression / translational elongation / ribosomal large subunit export from nucleus / 90S preribosome / G-protein alpha-subunit binding / positive regulation of protein kinase activity / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / regulation of translational fidelity / Ub-specific processing proteases / protein-RNA complex assembly / translation elongation factor activity / ribosomal subunit export from nucleus / translation regulator activity / translational termination / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of LSU-rRNA / cellular response to amino acid starvation / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rescue of stalled ribosome / ribosomal large subunit biogenesis / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / translational initiation / macroautophagy / protein kinase C binding / positive regulation of apoptotic signaling pathway / maintenance of translational fidelity / modification-dependent protein catabolic process / cytoplasmic stress granule / protein tag activity / rRNA processing / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / ribosome biogenesis / viral capsid / ribosome binding / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / negative regulation of translation / rRNA binding / ribosome / protein ubiquitination / structural constituent of ribosome / positive regulation of protein phosphorylation / G protein-coupled receptor signaling pathway / ribonucleoprotein complex / translation / negative regulation of gene expression / response to antibiotic / GTPase activity / mRNA binding / ubiquitin protein ligase binding / host cell nucleus / GTP binding / nucleolus / mitochondrion / RNA binding / zinc ion binding / nucleoplasm / nucleus / metal ion binding / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | Saccharomyces cerevisiae (パン酵母) / Escherichia coli (大腸菌) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 2.57 Å | |||||||||
データ登録者 | Milicevic N / Jenner L / Myasnikov A / Yusupov M / Yusupova G | |||||||||
資金援助 | フランス, 1件
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引用 | ジャーナル: Nature / 年: 2024 タイトル: mRNA reading frame maintenance during eukaryotic ribosome translocation. 著者: Nemanja Milicevic / Lasse Jenner / Alexander Myasnikov / Marat Yusupov / Gulnara Yusupova / 要旨: One of the most critical steps of protein synthesis is coupled translocation of messenger RNA (mRNA) and transfer RNAs (tRNAs) required to advance the mRNA reading frame by one codon. In ...One of the most critical steps of protein synthesis is coupled translocation of messenger RNA (mRNA) and transfer RNAs (tRNAs) required to advance the mRNA reading frame by one codon. In eukaryotes, translocation is accelerated and its fidelity is maintained by elongation factor 2 (eEF2). At present, only a few snapshots of eukaryotic ribosome translocation have been reported. Here we report ten high-resolution cryogenic-electron microscopy (cryo-EM) structures of the elongating eukaryotic ribosome bound to the full translocation module consisting of mRNA, peptidyl-tRNA and deacylated tRNA, seven of which also contained ribosome-bound, naturally modified eEF2. This study recapitulates mRNA-tRNA-growing peptide module progression through the ribosome, from the earliest states of eEF2 translocase accommodation until the very late stages of the process, and shows an intricate network of interactions preventing the slippage of the translational reading frame. We demonstrate how the accuracy of eukaryotic translocation relies on eukaryote-specific elements of the 80S ribosome, eEF2 and tRNAs. Our findings shed light on the mechanism of translation arrest by the anti-fungal eEF2-binding inhibitor, sordarin. We also propose that the sterically constrained environment imposed by diphthamide, a conserved eukaryotic posttranslational modification in eEF2, not only stabilizes correct Watson-Crick codon-anticodon interactions but may also uncover erroneous peptidyl-tRNA, and therefore contribute to higher accuracy of protein synthesis in eukaryotes. | |||||||||
履歴 |
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-構造の表示
添付画像 |
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-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_16634.map.gz | 413.8 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-16634-v30.xml emd-16634.xml | 109.6 KB 109.6 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_16634_fsc.xml | 19.7 KB | 表示 | FSCデータファイル |
画像 | emd_16634.png | 37.9 KB | ||
Filedesc metadata | emd-16634.cif.gz | 21.3 KB | ||
その他 | emd_16634_additional_1.map.gz emd_16634_additional_2.map.gz emd_16634_half_map_1.map.gz emd_16634_half_map_2.map.gz | 765.3 MB 765.6 MB 765 MB 764.7 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-16634 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-16634 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_16634_validation.pdf.gz | 1.3 MB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_16634_full_validation.pdf.gz | 1.3 MB | 表示 | |
XML形式データ | emd_16634_validation.xml.gz | 29 KB | 表示 | |
CIF形式データ | emd_16634_validation.cif.gz | 38.3 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-16634 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-16634 | HTTPS FTP |
-関連構造データ
関連構造データ | 8cg8MC 8ccsC 8cdlC 8cdrC 8cehC 8cf5C 8cgnC 8civC 8ckuC 8cmjC C: 同じ文献を引用 (文献) M: このマップから作成された原子モデル |
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類似構造データ | 類似検索 - 機能・相同性F&H 検索 |
-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_16634.map.gz / 形式: CCP4 / 大きさ: 824 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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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: 0.9 Å | ||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
-追加マップ: Half map A filtered to reported resolution
ファイル | emd_16634_additional_1.map | ||||||||||||
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注釈 | Half map A filtered to reported resolution | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-追加マップ: Half map B filtered to reported resolution
ファイル | emd_16634_additional_2.map | ||||||||||||
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注釈 | Half map B filtered to reported resolution | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: #2
ファイル | emd_16634_half_map_1.map | ||||||||||||
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投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: #1
ファイル | emd_16634_half_map_2.map | ||||||||||||
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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 : 80S Saccharomyces cerevisiae ribosome in complex with eEF2.sordar...
+超分子 #1: 80S Saccharomyces cerevisiae ribosome in complex with eEF2.sordar...
+分子 #1: 40S ribosomal protein S24-A
+分子 #2: 40S ribosomal protein S25-A
+分子 #3: 40S ribosomal protein S26
+分子 #4: 40S ribosomal protein S27-A
+分子 #5: 40S ribosomal protein S28-A
+分子 #6: HLJ1_G0030400.mRNA.1.CDS.1
+分子 #7: 40S ribosomal protein S30-A
+分子 #8: Guanine nucleotide-binding protein subunit beta-like protein
+分子 #9: Ubiquitin-40S ribosomal protein S31
+分子 #10: 60S ribosomal protein L16-A
+分子 #12: Elongation factor 2
+分子 #13: 60S ribosomal protein L17-A
+分子 #16: 60S ribosomal protein L18-A
+分子 #19: 60S ribosomal protein L19-A
+分子 #20: 60S acidic ribosomal protein P0
+分子 #22: 60S ribosomal protein L20-A
+分子 #23: 60S ribosomal protein L2-A
+分子 #24: 60S ribosomal protein L12-A
+分子 #25: 60S ribosomal protein L21-A
+分子 #26: 60S ribosomal protein L3
+分子 #27: 60S ribosomal protein L22-A
+分子 #28: 60S ribosomal protein L4-A
+分子 #29: 60S ribosomal protein L23-A
+分子 #30: 60S ribosomal protein L5
+分子 #31: 60S ribosomal protein L24-A
+分子 #32: 60S ribosomal protein L6-A
+分子 #33: 60S ribosomal protein L25
+分子 #34: 60S ribosomal protein L7-A
+分子 #35: 60S ribosomal protein L26-A
+分子 #36: 60S ribosomal protein L8-A
+分子 #37: 60S ribosomal protein L27-A
+分子 #38: RPL9A isoform 1
+分子 #39: 60S ribosomal protein L28
+分子 #40: 60S ribosomal protein L10
+分子 #41: 60S ribosomal protein L29
+分子 #42: 60S ribosomal protein L11-A
+分子 #43: 60S ribosomal protein L30
+分子 #44: 60S ribosomal protein L13-A
+分子 #45: 60S ribosomal protein L31-A
+分子 #46: 60S ribosomal protein L14-A
+分子 #47: Polypeptide
+分子 #48: 60S ribosomal protein L32
+分子 #49: 60S ribosomal protein L15-A
+分子 #50: 60S ribosomal protein L33-A
+分子 #51: 60S ribosomal protein L34-A
+分子 #52: 60S ribosomal protein L35-A
+分子 #53: 60S ribosomal protein L36-A
+分子 #54: 60S ribosomal protein L37-A
+分子 #55: 60S ribosomal protein L38
+分子 #56: 60S ribosomal protein L39
+分子 #57: Ubiquitin-60S ribosomal protein L40
+分子 #58: 60S ribosomal protein L41
+分子 #59: 60S ribosomal protein L42-A
+分子 #60: 60S ribosomal protein L43-A
+分子 #62: 40S ribosomal protein S0-A
+分子 #63: 40S ribosomal protein S1-A
+分子 #64: 40S ribosomal protein S2
+分子 #65: RPS3 isoform 1
+分子 #66: 40S ribosomal protein S4-A
+分子 #67: 40S ribosomal protein S5
+分子 #68: 40S ribosomal protein S6-A
+分子 #69: 40S ribosomal protein S7-A
+分子 #70: 40S ribosomal protein S8-B
+分子 #71: 40S ribosomal protein S9-A
+分子 #72: 40S ribosomal protein S10-A
+分子 #73: 40S ribosomal protein S11-A
+分子 #74: 40S ribosomal protein S13
+分子 #75: 40S ribosomal protein S14-A
+分子 #76: 40S ribosomal protein S15
+分子 #77: 40S ribosomal protein S16-A
+分子 #78: 40S ribosomal protein S17-A
+分子 #79: 40S ribosomal protein S18-A
+分子 #80: 40S ribosomal protein S19-A
+分子 #81: 40S ribosomal protein S20
+分子 #82: 40S ribosomal protein S21-A
+分子 #83: 40S ribosomal protein S22-A
+分子 #84: 40S ribosomal protein S23-A
+分子 #11: 25S ribosomal RNA
+分子 #14: 5S ribosomal RNA
+分子 #15: Transfer RNA Phe
+分子 #17: 5.8S ribosomal RNA
+分子 #18: Transfer RNA fMet
+分子 #21: Messenger RNA
+分子 #61: 18S ribosomal RNA
+分子 #85: ZINC ION
+分子 #86: MAGNESIUM ION
+分子 #87: SPERMIDINE
+分子 #88: POTASSIUM ION
+分子 #89: GUANOSINE-5'-TRIPHOSPHATE
+分子 #90: [1R-(1.ALPHA.,3A.BETA.,4.BETA.,4A.BETA.,7.BETA.,7A.ALPHA.,8A.BETA...
+分子 #91: water
-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 / 装置: FEI VITROBOT MARK IV |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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撮影 | フィルム・検出器のモデル: FEI FALCON IV (4k x 4k) 平均電子線量: 40.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | C2レンズ絞り径: 50.0 µm / 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): 1.0 µm / 最小 デフォーカス(公称値): 0.4 µm / 倍率(公称値): 270000 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |