+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-12632 | |||||||||
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タイトル | Mammalian pre-termination 80S ribosome with eRF1 and eRF3 bound by Blasticidin S. | |||||||||
マップデータ | ||||||||||
試料 |
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キーワード | Inhibitor 80S Termination Complex Blasticidin S Translation NMD / RIBOSOME | |||||||||
機能・相同性 | 機能・相同性情報 Protein hydroxylation / L13a-mediated translational silencing of Ceruloplasmin expression / SRP-dependent cotranslational protein targeting to membrane / Major pathway of rRNA processing in the nucleolus and cytosol / Formation of a pool of free 40S subunits / 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) / Translation initiation complex formation / Formation of the ternary complex, and subsequently, the 43S complex ...Protein hydroxylation / L13a-mediated translational silencing of Ceruloplasmin expression / SRP-dependent cotranslational protein targeting to membrane / Major pathway of rRNA processing in the nucleolus and cytosol / Formation of a pool of free 40S subunits / 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) / Translation initiation complex formation / Formation of the ternary complex, and subsequently, the 43S complex / Ribosomal scanning and start codon recognition / translation termination factor activity / Translesion synthesis by REV1 / Recognition of DNA damage by PCNA-containing replication complex / Translesion Synthesis by POLH / Downregulation of ERBB4 signaling / Spry regulation of FGF signaling / Downregulation of ERBB2:ERBB3 signaling / NOD1/2 Signaling Pathway / APC/C:Cdc20 mediated degradation of Cyclin B / APC-Cdc20 mediated degradation of Nek2A / EGFR downregulation / TCF dependent signaling in response to WNT / NRIF signals cell death from the nucleus / p75NTR recruits signalling complexes / NF-kB is activated and signals survival / Activated NOTCH1 Transmits Signal to the Nucleus / Downregulation of TGF-beta receptor signaling / TGF-beta receptor signaling in EMT (epithelial to mesenchymal transition) / Downregulation of SMAD2/3:SMAD4 transcriptional activity / SMAD2/SMAD3:SMAD4 heterotrimer regulates transcription / Senescence-Associated Secretory Phenotype (SASP) / Regulation of innate immune responses to cytosolic DNA / activated TAK1 mediates p38 MAPK activation / JNK (c-Jun kinases) phosphorylation and activation mediated by activated human TAK1 / Regulation of FZD by ubiquitination / N-glycan trimming in the ER and Calnexin/Calreticulin cycle / Regulation of TNFR1 signaling / TNFR1-induced NF-kappa-B signaling pathway / Translesion synthesis by POLK / Translesion synthesis by POLI / Regulation of necroptotic cell death / HDR through Homologous Recombination (HRR) / Josephin domain DUBs / Recruitment and ATM-mediated phosphorylation of repair and signaling proteins at DNA double strand breaks / Processing of DNA double-strand break ends / Formation of Incision Complex in GG-NER / Gap-filling DNA repair synthesis and ligation in GG-NER / Dual Incision in GG-NER / Fanconi Anemia Pathway / Regulation of TP53 Activity through Phosphorylation / Regulation of TP53 Degradation / Regulation of TP53 Activity through Methylation / Negative regulation of MET activity / PTK6 Regulates RTKs and Their Effectors AKT1 and DOK1 / Downregulation of ERBB2 signaling / E3 ubiquitin ligases ubiquitinate target proteins / Regulation of PTEN localization / ER Quality Control Compartment (ERQC) / Regulation of expression of SLITs and ROBOs / Interferon alpha/beta signaling / Endosomal Sorting Complex Required For Transport (ESCRT) / Activation of IRF3, IRF7 mediated by TBK1, IKKε (IKBKE) / IKK complex recruitment mediated by RIP1 / IRAK2 mediated activation of TAK1 complex / TRAF6-mediated induction of TAK1 complex within TLR4 complex / Alpha-protein kinase 1 signaling pathway / RAS processing / Pexophagy / Negative regulation of FLT3 / IRAK2 mediated activation of TAK1 complex upon TLR7/8 or 9 stimulation / Regulation of NF-kappa B signaling / Regulation of TBK1, IKKε (IKBKE)-mediated activation of IRF3, IRF7 / Regulation of pyruvate metabolism / SCF-beta-TrCP mediated degradation of Emi1 / Termination of translesion DNA synthesis / MAP3K8 (TPL2)-dependent MAPK1/3 activation / Ovarian tumor domain proteases / Cyclin D associated events in G1 / Negative regulators of DDX58/IFIH1 signaling / Regulation of BACH1 activity / Negative regulation of FGFR1 signaling / Negative regulation of FGFR2 signaling / Negative regulation of FGFR3 signaling / Negative regulation of FGFR4 signaling / Negative regulation of MAPK pathway / Synthesis of active ubiquitin: roles of E1 and E2 enzymes / Inactivation of CSF3 (G-CSF) signaling / Iron uptake and transport / Deactivation of the beta-catenin transactivating complex / Metalloprotease DUBs / Formation of TC-NER Pre-Incision Complex / Dual incision in TC-NER / Gap-filling DNA repair synthesis and ligation in TC-NER / Major pathway of rRNA processing in the nucleolus and cytosol / GTP hydrolysis and joining of the 60S ribosomal subunit / cytoplasmic translational termination / Autodegradation of Cdh1 by Cdh1:APC/C / APC/C:Cdc20 mediated degradation of Securin / APC/C:Cdh1 mediated degradation of Cdc20 and other APC/C:Cdh1 targeted proteins in late mitosis/early G1 類似検索 - 分子機能 | |||||||||
生物種 | Oryctolagus cuniculus (ウサギ) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 4.1 Å | |||||||||
データ登録者 | Powers KT / Yadav SKN | |||||||||
資金援助 | 英国, 1件
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引用 | ジャーナル: Nucleic Acids Res / 年: 2021 タイトル: Blasticidin S inhibits mammalian translation and enhances production of protein encoded by nonsense mRNA. 著者: Kyle T Powers / Flint Stevenson-Jones / Sathish K N Yadav / Beate Amthor / Joshua C Bufton / Ufuk Borucu / Dakang Shen / Jonas P Becker / Daria Lavysh / Matthias W Hentze / Andreas E Kulozik ...著者: Kyle T Powers / Flint Stevenson-Jones / Sathish K N Yadav / Beate Amthor / Joshua C Bufton / Ufuk Borucu / Dakang Shen / Jonas P Becker / Daria Lavysh / Matthias W Hentze / Andreas E Kulozik / Gabriele Neu-Yilik / Christiane Schaffitzel / 要旨: Deciphering translation is of paramount importance for the understanding of many diseases, and antibiotics played a pivotal role in this endeavour. Blasticidin S (BlaS) targets translation by binding ...Deciphering translation is of paramount importance for the understanding of many diseases, and antibiotics played a pivotal role in this endeavour. Blasticidin S (BlaS) targets translation by binding to the peptidyl transferase center of the large ribosomal subunit. Using biochemical, structural and cellular approaches, we show here that BlaS inhibits both translation elongation and termination in Mammalia. Bound to mammalian terminating ribosomes, BlaS distorts the 3'CCA tail of the P-site tRNA to a larger extent than previously reported for bacterial ribosomes, thus delaying both, peptide bond formation and peptidyl-tRNA hydrolysis. While BlaS does not inhibit stop codon recognition by the eukaryotic release factor 1 (eRF1), it interferes with eRF1's accommodation into the peptidyl transferase center and subsequent peptide release. In human cells, BlaS inhibits nonsense-mediated mRNA decay and, at subinhibitory concentrations, modulates translation dynamics at premature termination codons leading to enhanced protein production. | |||||||||
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_12632.map.gz | 15.2 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-12632-v30.xml emd-12632.xml | 103.7 KB 103.7 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_12632_fsc.xml | 10.7 KB | 表示 | FSCデータファイル |
画像 | emd_12632.png | 79.4 KB | ||
マスクデータ | emd_12632_msk_1.map | 103 MB | マスクマップ | |
Filedesc metadata | emd-12632.cif.gz | 21 KB | ||
その他 | emd_12632_half_map_1.map.gz emd_12632_half_map_2.map.gz | 80.9 MB 80.7 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-12632 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-12632 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_12632_validation.pdf.gz | 1.1 MB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_12632_full_validation.pdf.gz | 1.1 MB | 表示 | |
XML形式データ | emd_12632_validation.xml.gz | 17.7 KB | 表示 | |
CIF形式データ | emd_12632_validation.cif.gz | 23.4 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12632 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12632 | 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_12632.map.gz / 形式: CCP4 / 大きさ: 103 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: 1.35 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
-マスク #1
ファイル | emd_12632_msk_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: #2
ファイル | emd_12632_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_12632_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 : Mammalian pre-termination 80S ribosome with eRF1 and eRF3 bound b...
+超分子 #1: Mammalian pre-termination 80S ribosome with eRF1 and eRF3 bound b...
+分子 #1: 60S ribosomal protein L8
+分子 #2: uL4
+分子 #3: eL31
+分子 #4: 40S ribosomal protein S3
+分子 #5: S29
+分子 #6: 60S ribosomal protein L5
+分子 #7: Ribosomal protein L32
+分子 #8: 40S ribosomal protein S4
+分子 #9: 40S ribosomal protein S30
+分子 #10: eL29
+分子 #11: 60S ribosomal protein L6
+分子 #12: eL33
+分子 #13: Ribosomal protein S5
+分子 #14: 40S ribosomal protein S27a
+分子 #15: uL30
+分子 #16: 60S ribosomal protein L34
+分子 #17: 40S ribosomal protein S3a
+分子 #18: 40S ribosomal protein S6
+分子 #19: Epididymis tissue sperm binding protein Li 3a
+分子 #20: L7a
+分子 #21: uL29
+分子 #22: S7
+分子 #24: 40S ribosomal protein S27
+分子 #25: L9
+分子 #26: 60S ribosomal protein L36
+分子 #27: 40S ribosomal protein S8
+分子 #28: Eukaryotic peptide chain release factor subunit 1
+分子 #29: 60S ribosomal protein L10
+分子 #30: Ribosomal protein L37
+分子 #31: 40S ribosomal protein S9
+分子 #32: eRF3a
+分子 #33: 60S ribosomal protein L11
+分子 #34: L38
+分子 #35: 40S ribosomal protein S10
+分子 #36: 60S ribosomal protein L13
+分子 #37: eL39
+分子 #38: 40S ribosomal protein S11
+分子 #39: Ribosomal protein L14
+分子 #40: 60S RIBOSOMAL PROTEIN EL40
+分子 #41: 40S ribosomal protein S12
+分子 #42: Ribosomal protein L15
+分子 #43: 60s ribosomal protein l41
+分子 #44: ribosomal protein uS15
+分子 #45: 60S RIBOSOMAL PROTEIN UL13
+分子 #46: eL42
+分子 #47: uS11
+分子 #48: uL22
+分子 #49: ribosomal protein eL43
+分子 #50: 40S ribosomal protein uS19
+分子 #51: eL18
+分子 #52: eL28
+分子 #53: Ribosomal protein S16
+分子 #54: 60S ribosomal protein L19
+分子 #55: 60S acidic ribosomal protein P0
+分子 #56: 40S ribosomal protein S17
+分子 #57: 60S ribosomal protein L18a
+分子 #58: uL12
+分子 #59: 40S ribosomal protein uS13
+分子 #60: eL21
+分子 #61: eS19
+分子 #62: L22
+分子 #63: 40S ribosomal protein S20
+分子 #64: eL14
+分子 #65: 40S ribosomal protein S21
+分子 #66: 60S ribosomal protein L24-like protein
+分子 #68: Ribosomal protein S15a
+分子 #69: uL23
+分子 #71: 40S ribosomal protein uS12
+分子 #72: Ribosomal protein L26
+分子 #74: 40S ribosomal protein S24
+分子 #75: 60S ribosomal protein L27
+分子 #77: 40S ribosomal protein S25
+分子 #78: 60S ribosomal protein L27a
+分子 #79: 40S_SA_C domain-containing protein
+分子 #80: 40S ribosomal protein S26
+分子 #81: uL3
+分子 #82: eL30
+分子 #83: 40S ribosomal protein S2
+分子 #84: 40S ribosomal protein S28
+分子 #23: mRNA
+分子 #67: 28S Ribosomal RNA
+分子 #70: 5S Ribosomal RNA
+分子 #73: 5.8S Ribosomal RNA
+分子 #76: 18S Ribosomal RNA
+分子 #85: ZINC ION
+分子 #86: MAGNESIUM ION
+分子 #87: PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER
+分子 #88: BLASTICIDIN S
-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
濃度 | 0.63 mg/mL |
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緩衝液 | pH: 7.4 詳細: 50 mM HEPES pH 7.4 100 mM KOAc 5 mM Mg(Oac)2 1mM DTT |
グリッド | モデル: Quantifoil R2/2 / 材質: COPPER / メッシュ: 300 / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 時間: 30 sec. |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 70 % / チャンバー内温度: 288.15 K / 装置: LEICA EM GP 詳細: 30s sample incubation on grid followed by 1.1s blotting time.. |
詳細 | 165nM (OD260nm~10.5) |
-電子顕微鏡法
顕微鏡 | FEI TALOS ARCTICA |
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詳細 | Collected in super-resolution mode (0.675A pixel size) |
撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: SUPER-RESOLUTION / 撮影したグリッド数: 1 / 実像数: 3500 / 平均露光時間: 8.0 sec. / 平均電子線量: 41.92 e/Å2 |
電子線 | 加速電圧: 200 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | C2レンズ絞り径: 100.0 µm / 最大 デフォーカス(補正後): 2.0 µm / 最小 デフォーカス(補正後): 0.4 µm / 倍率(補正後): 79000 / 照射モード: OTHER / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm |
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: Talos Arctica / 画像提供: FEI Company |