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基本情報
登録情報 | データベース: EMDB / ID: EMD-8190 | |||||||||
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タイトル | m48S late-stage initiation complex, purified from rabbit reticulocytes lysates, displaying eIF2 ternary complex and eIF3 i and g subunits relocated to the intersubunit face | |||||||||
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![]() | eukaryotic translation initiation / ribosome / eIF3 peripheral subunits / cryo-EM / translation | |||||||||
機能・相同性 | ![]() viral translational termination-reinitiation / eukaryotic translation initiation factor 3 complex, eIF3m / eukaryotic translation initiation factor 2 complex / eukaryotic translation initiation factor 3 complex / formation of cytoplasmic translation initiation complex / eukaryotic 43S preinitiation complex / formation of translation preinitiation complex / eukaryotic 48S preinitiation complex / ribosomal subunit / protein-synthesizing GTPase ...viral translational termination-reinitiation / eukaryotic translation initiation factor 3 complex, eIF3m / eukaryotic translation initiation factor 2 complex / eukaryotic translation initiation factor 3 complex / formation of cytoplasmic translation initiation complex / eukaryotic 43S preinitiation complex / formation of translation preinitiation complex / eukaryotic 48S preinitiation complex / ribosomal subunit / protein-synthesizing GTPase / Formation of the ternary complex, and subsequently, the 43S complex / laminin receptor activity / Ribosomal scanning and start codon recognition / Translation initiation complex formation / positive regulation of signal transduction by p53 class mediator / Formation of a pool of free 40S subunits / ubiquitin ligase inhibitor activity / phagocytic cup / 90S preribosome / GTP hydrolysis and joining of the 60S ribosomal subunit / L13a-mediated translational silencing of Ceruloplasmin expression / regulation of translational fidelity / ribosomal small subunit export from nucleus / rough endoplasmic reticulum / laminin binding / translation regulator activity / gastrulation / MDM2/MDM4 family protein binding / cytosolic ribosome / translation initiation factor activity / class I DNA-(apurinic or apyrimidinic site) endonuclease activity / DNA-(apurinic or apyrimidinic site) lyase / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / 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 / translational initiation / small-subunit processome / spindle / cytoplasmic stress granule / rRNA processing / rhythmic process / positive regulation of canonical Wnt signaling pathway / regulation of translation / ribosome binding / virus receptor activity / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / cytosolic small ribosomal subunit / small ribosomal subunit rRNA binding / perikaryon / cytosolic large ribosomal subunit / cytoplasmic translation / cell differentiation / tRNA binding / mitochondrial inner membrane / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / cell division / DNA repair / GTPase activity / mRNA binding / apoptotic process / synapse / centrosome / dendrite / GTP binding / nucleolus / perinuclear region of cytoplasm / Golgi apparatus / DNA binding / RNA binding / zinc ion binding / nucleus / membrane / plasma membrane / cytosol / cytoplasm 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: 5.8 Å | |||||||||
![]() | Simonetti A / Brito Querido J / Myasnikov AG / Mancera-Martinez E / Renaud A / Kuhn L / Hashem Y | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: eIF3 Peripheral Subunits Rearrangement after mRNA Binding and Start-Codon Recognition. 著者: Angelita Simonetti / Jailson Brito Querido / Alexander G Myasnikov / Eder Mancera-Martinez / Adeline Renaud / Lauriane Kuhn / Yaser Hashem / ![]() 要旨: mRNA translation initiation in eukaryotes requires the cooperation of a dozen eukaryotic initiation factors (eIFs) forming several complexes, which leads to mRNA attachment to the small ribosomal ...mRNA translation initiation in eukaryotes requires the cooperation of a dozen eukaryotic initiation factors (eIFs) forming several complexes, which leads to mRNA attachment to the small ribosomal 40S subunit, mRNA scanning for start codon, and accommodation of initiator tRNA at the 40S P site. eIF3, composed of 13 subunits, 8 core (a, c, e, f, h, l, k, and m) and 5 peripheral (b, d, g, i, and j), plays a central role during this process. Here we report a cryo-electron microscopy structure of a mammalian 48S initiation complex at 5.8 Å resolution. It shows the relocation of subunits eIF3i and eIF3g to the 40S intersubunit face on the GTPase binding site, at a late stage in initiation. On the basis of a previous study, we demonstrate the relocation of eIF3b to the 40S intersubunit face, binding below the eIF2-Met-tRNAi(Met) ternary complex upon mRNA attachment. Our analysis reveals the deep rearrangement of eIF3 and unravels the molecular mechanism underlying eIF3 function in mRNA scanning and timing of ribosomal subunit joining. | |||||||||
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構造ビューア | EMマップ: ![]() ![]() ![]() |
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マップデータ | ![]() | 28.6 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 66.9 KB 66.9 KB | 表示 表示 | ![]() |
画像 | ![]() | 54.3 KB | ||
Filedesc metadata | ![]() | 13.3 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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EMDBのページ | ![]() ![]() |
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注釈 | None | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 2.2 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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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
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試料の構成要素
+全体 : m48S late-stage initiation complex, purified from rabbit reticulo...
+超分子 #1: m48S late-stage initiation complex, purified from rabbit reticulo...
+分子 #1: tRNA
+分子 #2: 18S ribosomal RNA
+分子 #3: mRNA
+分子 #4: Eukaryotic translation initiation factor 2 subunit 1
+分子 #5: ribosomal protein uS17
+分子 #6: ribosomal protein uS9
+分子 #7: 40S ribosomal protein S4
+分子 #8: ribosomal protein uS14
+分子 #9: Ribosomal protein S9 (Predicted)
+分子 #10: ribosomal protein uS13
+分子 #11: Eukaryotic translation initiation factor 3 subunit G
+分子 #12: Eukaryotic translation initiation factor 3 subunit G
+分子 #13: ribosomal protein uS12
+分子 #14: ribosomal protein eS19
+分子 #15: eukaryotic initiation factor 2 Gamma subunit (eIF2-Gamma)
+分子 #16: Eukaryotic translation initiation factor 3 subunit I
+分子 #17: ribosomal protein uS7
+分子 #18: ribosomal protein eS30
+分子 #19: ribosomal protein eS25
+分子 #20: ribosomal protein eS7
+分子 #21: 40S ribosomal protein S27
+分子 #22: ribosomal protein uS15
+分子 #23: ribosomal protein uS8
+分子 #24: 40S ribosomal protein S21
+分子 #25: ribosomal protein uS5
+分子 #26: eukaryotic initiation factor 2 subunit Beta (eIF2-Beta)
+分子 #27: ribosomal protein eS17
+分子 #28: ribosomal protein uS2
+分子 #29: ribosomal protein uS3
+分子 #30: ribosomal protein uS10
+分子 #31: ribosomal protein eS1
+分子 #32: ribosomal protein uS11
+分子 #33: ribosomal protein eS26
+分子 #34: ribosomal protein eS28
+分子 #35: ribosomal protein RACK1
+分子 #36: ribosomal protein uS19
+分子 #37: 40S ribosomal protein S8
+分子 #38: ribosomal protein eS31
+分子 #39: 40S ribosomal protein S6
+分子 #40: 40S ribosomal protein S12
+分子 #41: 40S ribosomal protein S24
+分子 #42: ribosomal protein eS10
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 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 | 0.5 mg/mL |
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緩衝液 | pH: 7.6 |
グリッド | モデル: Quantifoil / 材質: COPPER / メッシュ: 400 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: HOLEY / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 時間: 25 sec. / 前処理 - 雰囲気: AIR |
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 |
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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 II (4k x 4k) 検出モード: INTEGRATING / デジタル化 - 画像ごとのフレーム数: 2-8 / 撮影したグリッド数: 1 / 実像数: 5700 / 平均露光時間: 1.5 sec. / 平均電子線量: 24.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | C2レンズ絞り径: 100.0 µm / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 0.01 mm / 最大 デフォーカス(公称値): 4.5 µm / 最小 デフォーカス(公称値): 0.8 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: Titan Krios / 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 | PDB ID: Chain - Source name: PDB / Chain - Initial model type: experimental model |
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精密化 | プロトコル: FLEXIBLE FIT |
得られたモデル | ![]() PDB-5k0y: |