+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-0058 | |||||||||
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タイトル | Cryo-EM map of a partial yeast 48S preinitiation complex in its closed conformation obtained by focused classification using masks for eIF3b-eIF3i. | |||||||||
マップデータ | For optimal visualization of eIF2, and of eIF3c, eIF3b and eIf3a c-term at the subunit interface, gauss-filter the map by 1.34 and display it at 0.035 | |||||||||
試料 |
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キーワード | ribosome / translation / initiation factors / 40S / eIF1A / eIF3 / eIF2 / tRNAi / 48S PIC / small ribosome subunit | |||||||||
機能・相同性 | 機能・相同性情報 formation of translation initiation ternary complex / Recycling of eIF2:GDP / Cellular response to mitochondrial stress / eukaryotic translation initiation factor 3 complex, eIF3e / ABC-family proteins mediated transport / eukaryotic translation initiation factor 3 complex, eIF3m / methionyl-initiator methionine tRNA binding / incipient cellular bud site / translation reinitiation / eukaryotic translation initiation factor 2 complex ...formation of translation initiation ternary complex / Recycling of eIF2:GDP / Cellular response to mitochondrial stress / eukaryotic translation initiation factor 3 complex, eIF3e / ABC-family proteins mediated transport / eukaryotic translation initiation factor 3 complex, eIF3m / methionyl-initiator methionine tRNA binding / incipient cellular bud site / translation reinitiation / eukaryotic translation initiation factor 2 complex / eukaryotic translation initiation factor 3 complex / formation of cytoplasmic translation initiation complex / multi-eIF complex / cytoplasmic translational initiation / protein-synthesizing GTPase / eukaryotic 43S preinitiation complex / formation of translation preinitiation complex / eukaryotic 48S preinitiation complex / positive regulation of translational fidelity / Formation of the ternary complex, and subsequently, the 43S complex / Translation initiation complex formation / Ribosomal scanning and start codon recognition / Formation of a pool of free 40S subunits / L13a-mediated translational silencing of Ceruloplasmin expression / ribosomal small subunit binding / 90S preribosome / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / regulation of translational fidelity / 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) / translation initiation factor binding / translation initiation factor activity / rescue of stalled ribosome / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / translational initiation / protein kinase C binding / maintenance of translational fidelity / modification-dependent protein catabolic process / cytoplasmic stress granule / protein tag activity / rRNA processing / ribosomal small subunit biogenesis / double-stranded RNA binding / small ribosomal subunit rRNA binding / ribosome binding / ribosomal small subunit assembly / small ribosomal subunit / cytosolic small ribosomal subunit / cytoplasmic translation / cytosolic large ribosomal subunit / rRNA binding / ribosome / structural constituent of ribosome / protein ubiquitination / translation / ribonucleoprotein complex / positive regulation of protein phosphorylation / GTPase activity / mRNA binding / ubiquitin protein ligase binding / nucleolus / GTP binding / protein kinase binding / RNA binding / zinc ion binding / identical protein binding / nucleus / metal ion binding / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (パン酵母) / Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37) (酵母) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 5.75 Å | |||||||||
データ登録者 | Llacer JL / Hussain T | |||||||||
資金援助 | 英国, 2件
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引用 | ジャーナル: Mol Cell / 年: 2015 タイトル: Conformational Differences between Open and Closed States of the Eukaryotic Translation Initiation Complex. 著者: Jose L Llácer / Tanweer Hussain / Laura Marler / Colin Echeverría Aitken / Anil Thakur / Jon R Lorsch / Alan G Hinnebusch / V Ramakrishnan / 要旨: Translation initiation in eukaryotes begins with the formation of a pre-initiation complex (PIC) containing the 40S ribosomal subunit, eIF1, eIF1A, eIF3, ternary complex (eIF2-GTP-Met-tRNAi), and ...Translation initiation in eukaryotes begins with the formation of a pre-initiation complex (PIC) containing the 40S ribosomal subunit, eIF1, eIF1A, eIF3, ternary complex (eIF2-GTP-Met-tRNAi), and eIF5. The PIC, in an open conformation, attaches to the 5' end of the mRNA and scans to locate the start codon, whereupon it closes to arrest scanning. We present single particle cryo-electron microscopy (cryo-EM) reconstructions of 48S PICs from yeast in these open and closed states, at 6.0 Å and 4.9 Å, respectively. These reconstructions show eIF2β as well as a configuration of eIF3 that appears to encircle the 40S, occupying part of the subunit interface. Comparison of the complexes reveals a large conformational change in the 40S head from an open mRNA latch conformation to a closed one that constricts the mRNA entry channel and narrows the P site to enclose tRNAi, thus elucidating key events in start codon recognition. | |||||||||
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_0058.map.gz | 96.4 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-0058-v30.xml emd-0058.xml | 83.2 KB 83.2 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_0058.png | 75.3 KB | ||
Filedesc metadata | emd-0058.cif.gz | 15.6 KB | ||
その他 | emd_0058_half_map_1.map.gz emd_0058_half_map_2.map.gz | 80.8 MB 80.8 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-0058 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-0058 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_0058_validation.pdf.gz | 1 MB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_0058_full_validation.pdf.gz | 1 MB | 表示 | |
XML形式データ | emd_0058_validation.xml.gz | 13.1 KB | 表示 | |
CIF形式データ | emd_0058_validation.cif.gz | 15.4 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-0058 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-0058 | 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_0058.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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注釈 | For optimal visualization of eIF2, and of eIF3c, eIF3b and eIf3a c-term at the subunit interface, gauss-filter the map by 1.34 and display it at 0.035 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.34 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
-ハーフマップ: #1
ファイル | emd_0058_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: #2
ファイル | emd_0058_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 : Structure of a partial yeast 48S preinitiation complex in closed ...
+超分子 #1: Structure of a partial yeast 48S preinitiation complex in closed ...
+超分子 #3: tRNA
+超分子 #4: Initiation factors
+超分子 #5: mRNA
+超分子 #6: Initiation factors
+超分子 #2: Ribosome
+分子 #1: 18S rRNA (1798-MER)
+分子 #20: mRNA
+分子 #28: tRNAi (75-MER)
+分子 #2: KLLA0F09812p
+分子 #3: KLLA0D08305p
+分子 #4: KLLA0D10659p
+分子 #5: KLLA0B08173p
+分子 #6: 40S ribosomal protein S12
+分子 #7: KLLA0F07843p
+分子 #8: 40S ribosomal protein S16
+分子 #9: KLLA0B01474p
+分子 #10: KLLA0B01562p
+分子 #11: KLLA0A07194p
+分子 #12: KLLA0F25542p
+分子 #13: KLLA0B06182p
+分子 #14: 40S ribosomal protein S28
+分子 #15: Ubiquitin-40S ribosomal protein S27a
+分子 #16: KLLA0E12277p
+分子 #17: 40S ribosomal protein S29
+分子 #18: 40S ribosomal protein S30
+分子 #19: 60S ribosomal protein L41-A
+分子 #21: Eukaryotic translation initiation factor 1A
+分子 #22: Eukaryotic translation initiation factor eIF-1
+分子 #23: Eukaryotic translation initiation factor 3 subunit A,eIF3a
+分子 #24: Eukaryotic translation initiation factor 3 subunit C
+分子 #25: Eukaryotic translation initiation factor 2 subunit gamma
+分子 #26: Eukaryotic translation initiation factor 2 subunit beta
+分子 #27: Eukaryotic translation initiation factor 2 subunit alpha
+分子 #29: 40S ribosomal protein S0
+分子 #30: 40S ribosomal protein S1
+分子 #31: 40S ribosomal protein S4
+分子 #32: 40S ribosomal protein S6
+分子 #33: 40S ribosomal protein S7
+分子 #34: 40S ribosomal protein S8
+分子 #35: KLLA0E23673p
+分子 #36: KLLA0A10483p
+分子 #37: KLLA0F18040p
+分子 #38: 40S ribosomal protein S14
+分子 #39: 40S ribosomal protein S21
+分子 #40: 40S ribosomal protein S22
+分子 #41: 40S ribosomal protein S24
+分子 #42: RPS23
+分子 #43: 40S ribosomal protein S26
+分子 #44: 40S ribosomal protein S27
+分子 #45: Eukaryotic translation initiation factor 3 subunit B
+分子 #46: Eukaryotic translation initiation factor 3 subunit I
+分子 #47: Eukaryotic translation initiation factor 3 subunit G
+分子 #48: MAGNESIUM ION
+分子 #49: ZINC ION
+分子 #50: PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER
+分子 #51: METHIONINE
-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 | 0.18 mg/mL | ||||||||||||||
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緩衝液 | pH: 6.5 構成要素:
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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 I |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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温度 | 最低: 90.0 K / 最高: 100.0 K |
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 / 撮影したグリッド数: 3 / 実像数: 5500 / 平均露光時間: 1.1 sec. / 平均電子線量: 32.0 e/Å2 詳細: Images were collected in movie-mode at 32 frames per second |
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: 70.0 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): 3.5 µm / 最小 デフォーカス(公称値): 1.5 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: Titan Krios / 画像提供: FEI Company |
+画像解析
-原子モデル構築 1
精密化 | 空間: RECIPROCAL / プロトコル: OTHER / 当てはまり具合の基準: FSC |
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得られたモデル | PDB-6gsn: |