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データを開く
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基本情報
| 登録情報 | ![]() | ||||||||||||
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| タイトル | Late-stage 48S Initiation Complex (LS48S IC) guided by the trans-RNA | ||||||||||||
マップデータ | main map of the mammalian late-stage 48S initiation complex | ||||||||||||
試料 |
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キーワード | 48S late-stage initiation complex / Programmable mRNA translation initiation / trans-RNA / activation of mRNA translation / TRANSLATION | ||||||||||||
| 機能・相同性 | 機能・相同性情報regulation of macromolecule metabolic process / Translation initiation complex formation / Formation of the ternary complex, and subsequently, the 43S complex / Ribosomal scanning and start codon recognition / eukaryotic initiation factor eIF2 binding / Major pathway of rRNA processing in the nucleolus and cytosol / GTP hydrolysis and joining of the 60S ribosomal subunit / L13a-mediated translational silencing of Ceruloplasmin expression / SRP-dependent cotranslational protein targeting to membrane / Formation of a pool of free 40S subunits ...regulation of macromolecule metabolic process / Translation initiation complex formation / Formation of the ternary complex, and subsequently, the 43S complex / Ribosomal scanning and start codon recognition / eukaryotic initiation factor eIF2 binding / Major pathway of rRNA processing in the nucleolus and cytosol / GTP hydrolysis and joining of the 60S ribosomal subunit / L13a-mediated translational silencing of Ceruloplasmin expression / SRP-dependent cotranslational protein targeting to membrane / Formation of a pool of free 40S subunits / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / eukaryotic translation initiation factor 2 complex / formation of cytoplasmic translation initiation complex / eukaryotic 48S preinitiation complex / cellular response to chemical stress / GDP-dissociation inhibitor activity / laminin receptor activity / 90S preribosome / ubiquitin ligase inhibitor activity / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / positive regulation of signal transduction by p53 class mediator / phagocytic cup / translation regulator activity / 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) / rough endoplasmic reticulum / ribosomal small subunit export from nucleus / laminin binding / translation initiation factor activity / gastrulation / MDM2/MDM4 family protein binding / cytosolic ribosome / class I DNA-(apurinic or apyrimidinic site) endonuclease activity / response to endoplasmic reticulum stress / DNA-(apurinic or apyrimidinic site) lyase / GTPase activator activity / positive regulation of apoptotic signaling pathway / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / spindle / cytoplasmic stress granule / rRNA processing / regulation of translation / rhythmic process / positive regulation of canonical Wnt signaling pathway / ribosomal small subunit assembly / virus receptor activity / ribosome binding / ribosomal small subunit biogenesis / small ribosomal subunit / small ribosomal subunit rRNA binding / cytosolic small ribosomal subunit / perikaryon / cell differentiation / cytoplasmic translation / tRNA binding / mitochondrial inner membrane / postsynaptic density / rRNA binding / structural constituent of ribosome / ribosome / translation / ribonucleoprotein complex / cell division / DNA repair / mRNA binding / apoptotic process / centrosome / synapse / dendrite / GTP binding / nucleolus / perinuclear region of cytoplasm / Golgi apparatus / ATP hydrolysis activity / DNA binding / RNA binding / zinc ion binding / ATP binding / membrane / nucleus / plasma membrane / cytoplasm / cytosol 類似検索 - 分子機能 | ||||||||||||
| 生物種 | ![]() | ||||||||||||
| 手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 2.9 Å | ||||||||||||
データ登録者 | Nguyen TT / Hashem Y / Rocha REO / Boissier F / Qian SB / Jia L / Uematsu S / Gu Y / Shi S | ||||||||||||
| 資金援助 | 米国, European Union, 3件
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引用 | ジャーナル: Nat Biotechnol / 年: 2025タイトル: Programmable initiation of mRNA translation by trans-RNA. 著者: Longfei Jia / Tan-Trung Nguyen / Saori Uematsu / Yifei Gu / Shengcho Shi / Yaser Hashem / Shu-Bing Qian / ![]() 要旨: Several approaches exist to silence genes, but few tools are available to activate individual mRNAs for translation inside cells. Guiding ribosomes to specific start codons without altering the ...Several approaches exist to silence genes, but few tools are available to activate individual mRNAs for translation inside cells. Guiding ribosomes to specific start codons without altering the original sequence remains a formidable task. Here we design capped trans-RNAs capable of directing ribosomes to specific initiation sites on individual mRNAs when the trans-cap is positioned near the target start codon. Structural and biochemical data suggest that the capped trans-RNA facilitates ribosome loading and scanning on the target mRNA through a synergistic mechanism involving alternative cap recognition. The trans-RNA also acts independently of the cap on the target mRNA, enabling translation of circular RNAs lacking internal ribosome entry sites. We apply trans-RNAs in vivo to achieve programmable alternative translation of endogenous genes in mouse liver. Finally, we provide the evidence for the existence of natural transcripts that, similarly to exogenous trans-RNAs, activate translation of endogenous mRNAs. | ||||||||||||
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構造の表示
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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_51903.map.gz | 450.4 MB | EMDBマップデータ形式 | |
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| ヘッダ (付随情報) | emd-51903-v30.xml emd-51903.xml | 70.7 KB 70.7 KB | 表示 表示 | EMDBヘッダ |
| FSC (解像度算出) | emd_51903_fsc.xml | 16.5 KB | 表示 | FSCデータファイル |
| 画像 | emd_51903.png | 71.5 KB | ||
| Filedesc metadata | emd-51903.cif.gz | 15.3 KB | ||
| その他 | emd_51903_additional_1.map.gz emd_51903_half_map_1.map.gz emd_51903_half_map_2.map.gz | 397.8 MB 442.5 MB 442.5 MB | ||
| アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-51903 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-51903 | HTTPS FTP |
-関連構造データ
| 関連構造データ | ![]() 9h6yMC ![]() 9h74C M: このマップから作成された原子モデル C: 同じ文献を引用 ( |
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| 類似構造データ | 類似検索 - 機能・相同性 F&H 検索 |
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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 |
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マップ
| ファイル | ダウンロード / ファイル: emd_51903.map.gz / 形式: CCP4 / 大きさ: 476.8 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| 注釈 | main map of the mammalian late-stage 48S initiation complex | ||||||||||||||||||||||||||||||||||||
| 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: 0.93 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: EMhanced map of the mammalian late-stage 48S initiation complex
| ファイル | emd_51903_additional_1.map | ||||||||||||
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| 注釈 | EMhanced map of the mammalian late-stage 48S initiation complex | ||||||||||||
| 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 B of the mammalian late-stage 48S initiation complex
| ファイル | emd_51903_half_map_1.map | ||||||||||||
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| 注釈 | half map B of the mammalian late-stage 48S initiation complex | ||||||||||||
| 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 A of the mammalian late-stage 48S initiation complex
| ファイル | emd_51903_half_map_2.map | ||||||||||||
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| 注釈 | half map A of the mammalian late-stage 48S initiation complex | ||||||||||||
| 投影像・断面図 |
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| 密度ヒストグラム |
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試料の構成要素
+全体 : Late-stage 48S Initiation Complex (LS48S IC) guided by the trans-...
+超分子 #1: Late-stage 48S Initiation Complex (LS48S IC) guided by the trans-...
+分子 #1: Initiator methionylated tRNA
+分子 #2: 18S ribosomal RNA
+分子 #3: Linear mRNA
+分子 #4: Eukaryotic translation initiation factor 2 subunit 1
+分子 #5: Eukaryotic translation initiation factor 2 subunit 3
+分子 #6: Small ribosomal subunit protein uS2
+分子 #7: Small ribosomal subunit protein eS1
+分子 #8: Small ribosomal subunit protein uS5
+分子 #9: Small ribosomal subunit protein uS3
+分子 #10: Small ribosomal subunit protein eS4
+分子 #11: Small ribosomal subunit protein uS7
+分子 #12: Small ribosomal subunit protein eS6
+分子 #13: Small ribosomal subunit protein eS7
+分子 #14: Small ribosomal subunit protein eS8
+分子 #15: Small ribosomal subunit protein uS4
+分子 #16: Small ribosomal subunit protein eS10
+分子 #17: Small ribosomal subunit protein uS17
+分子 #18: Small ribosomal subunit protein eS12
+分子 #19: Small ribosomal subunit protein uS15
+分子 #20: Small ribosomal subunit protein uS11
+分子 #21: Small ribosomal subunit protein uS19
+分子 #22: Small ribosomal subunit protein uS9
+分子 #23: Small ribosomal subunit protein eS17
+分子 #24: Small ribosomal subunit protein uS13
+分子 #25: Small ribosomal subunit protein eS19
+分子 #26: Small ribosomal subunit protein uS10
+分子 #27: Small ribosomal subunit protein eS21
+分子 #28: Small ribosomal subunit protein uS8
+分子 #29: Small ribosomal subunit protein uS12
+分子 #30: 40S ribosomal protein S24
+分子 #31: Small ribosomal subunit protein eS26
+分子 #32: Small ribosomal subunit protein eS27
+分子 #33: Small ribosomal subunit protein eS28
+分子 #34: Small ribosomal subunit protein uS14
+分子 #35: Ubiquitin-ribosomal protein eS31 fusion protein
+分子 #36: Small ribosomal subunit protein RACK1
+分子 #37: Ubiquitin-like FUBI-ribosomal protein eS30 fusion protein
+分子 #38: Eukaryotic translation initiation factor 4C
+分子 #39: ATP binding cassette subfamily E member 1
+分子 #40: Small ribosomal subunit protein eS32
+分子 #41: Small ribosomal subunit protein eS25
+分子 #42: Eukaryotic translation initiation factor 5
+分子 #43: IRON/SULFUR CLUSTER
-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 |
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試料調製
| 濃度 | 0.4 mg/mL | ||||||||||||||||||
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| 緩衝液 | pH: 7.4 構成要素:
詳細: 10 mM HEPES-KOH pH 7.4, 5 mM Mg(OAc)2, 50 mM KOAc, 10 mM NH4Cl, 2 mM DTT and Roche cOmplete protease inhibitor cocktail | ||||||||||||||||||
| グリッド | モデル: Quantifoil R2/2 / 材質: COPPER / メッシュ: 300 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: HOLEY / 支持フィルム - Film thickness: 2 / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 時間: 20 sec. / 詳細: glow-discharged at 2.5 mA for 20 s | ||||||||||||||||||
| 凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277 K / 装置: FEI VITROBOT MARK IV 詳細: The sample was incubated on the grid for 30 s and then blotted with filter paper from both sides for 1.5 s with the blot force 5. | ||||||||||||||||||
| 詳細 | late-stage 48S initiation translation complex was dissolved in 20 microlitre buffer containing 10 mM HEPES-KOH pH 7.4, 5 mM Mg(OAc)2, 50 mM KOAc, 10 mM NH4Cl, 2 mM DTT and Roche cOmplete protease inhibitor cocktail |
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電子顕微鏡法
| 顕微鏡 | FEI TALOS ARCTICA |
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| 温度 | 最低: 90.0 K / 最高: 95.0 K |
| 撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: COUNTING / デジタル化 - サイズ - 横: 4096 pixel / デジタル化 - サイズ - 縦: 4096 pixel / デジタル化 - 画像ごとのフレーム数: 1-40 / 撮影したグリッド数: 1 / 実像数: 9524 / 平均露光時間: 6.0 sec. / 平均電子線量: 50.0 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: 30.0 µm / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm / 最大 デフォーカス(公称値): 2.5 µm / 最小 デフォーカス(公称値): 0.5 µm / 倍率(公称値): 59000 |
| 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 |
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画像解析
-原子モデル構築 1
| 初期モデル |
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| 詳細 | The atomic model of LS48S IC complex (PDB: 8P09) was initially rigid body fitted in density using ChimeraX and further model building was done for mRNA, eIF5 N-terminal domain, eIF2gamma, eIF3d and eIF3 octamer in ChimeraX. Densities for eIF5 N-terminal domain, eIF2gamma, eIF3d and eIF3 octamer were observed in the map and therefore were included in the model. The 3D structure of the eIF5-N terminal domain was generated by Alphafold2 (PDB: G1TGW1). It was then accommodated into the model by aligning structurally to its counterparts in the yeast atomic model of the py48S-eIF5N pre-initiation complex (PDB: 6FYX). IF3d and eIF3 octamer were taken from the atomic model of LS48S IC complex with eIF3 proteins (PDB: 6YAM) and placed into density by rigid-body fitting. The atomic models with and without eIF3 proteins were inspected visually and performed stereochemical, torsional, angle refinement and optimization by using ISOLDE v1.6. Figures featuring cryo-EM densities and atomic models were visualized by using ChimeraX. | |||||||||||||||
| 精密化 | プロトコル: RIGID BODY FIT | |||||||||||||||
| 得られたモデル | ![]() PDB-9h6y: |
ムービー
コントローラー
万見について




キーワード
データ登録者
米国, European Union, 3件
引用
























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FIELD EMISSION GUN





