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データを開く
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基本情報
| 登録情報 | データベース: PDB / ID: 6fec | |||||||||
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| タイトル | Human cap-dependent 48S pre-initiation complex | |||||||||
要素 |
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キーワード | RIBOSOME / Translation initiation / 48S complex / capped mRNA / initiation factor 4B / start codon recognition | |||||||||
| 機能・相同性 | 機能・相同性情報eukaryotic translation initiation factor 4F complex assembly / RNA strand-exchange activity / positive regulation of mRNA binding / viral translational termination-reinitiation / Cellular response to mitochondrial stress / eukaryotic translation initiation factor 3 complex, eIF3e / Response of EIF2AK1 (HRI) to heme deficiency / Recycling of eIF2:GDP / eukaryotic translation initiation factor 3 complex, eIF3m / Activation of the mRNA upon binding of the cap-binding complex and eIFs, and subsequent binding to 43S ...eukaryotic translation initiation factor 4F complex assembly / RNA strand-exchange activity / positive regulation of mRNA binding / viral translational termination-reinitiation / Cellular response to mitochondrial stress / eukaryotic translation initiation factor 3 complex, eIF3e / Response of EIF2AK1 (HRI) to heme deficiency / Recycling of eIF2:GDP / eukaryotic translation initiation factor 3 complex, eIF3m / Activation of the mRNA upon binding of the cap-binding complex and eIFs, and subsequent binding to 43S / methionyl-initiator methionine tRNA binding / RNA strand annealing activity / PERK regulates gene expression / eukaryotic translation initiation factor 2 complex / eukaryotic translation initiation factor 3 complex / eukaryotic translation initiation factor 4F complex / formation of cytoplasmic translation initiation complex / Z-decay: degradation of maternal mRNAs by zygotically expressed factors / cytoplasmic translational initiation / eukaryotic 43S preinitiation complex / translation factor activity, RNA binding / formation of translation preinitiation complex / Deadenylation of mRNA / eukaryotic 48S preinitiation complex / M-decay: degradation of maternal mRNAs by maternally stored factors / negative regulation of endoplasmic reticulum unfolded protein response / oxidized pyrimidine DNA binding / response to TNF agonist / positive regulation of base-excision repair / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage / positive regulation of gastrulation / protein tyrosine kinase inhibitor activity / positive regulation of endodeoxyribonuclease activity / IRE1-RACK1-PP2A complex / positive regulation of Golgi to plasma membrane protein transport / TNFR1-mediated ceramide production / negative regulation of DNA repair / negative regulation of RNA splicing / protein-synthesizing GTPase / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / supercoiled DNA binding / neural crest cell differentiation / regulation of translational initiation / NF-kappaB complex / cysteine-type endopeptidase activator activity involved in apoptotic process / oxidized purine DNA binding / negative regulation of intrinsic apoptotic signaling pathway in response to hydrogen peroxide / regulation of establishment of cell polarity / negative regulation of bicellular tight junction assembly / ubiquitin-like protein conjugating enzyme binding / negative regulation of phagocytosis / rRNA modification in the nucleus and cytosol / erythrocyte homeostasis / Formation of the ternary complex, and subsequently, the 43S complex / cytoplasmic side of rough endoplasmic reticulum membrane / laminin receptor activity / negative regulation of ubiquitin protein ligase activity / protein kinase A binding / ion channel inhibitor activity / Ribosomal scanning and start codon recognition / pigmentation / Translation initiation complex formation / positive regulation of mitochondrial depolarization / positive regulation of T cell receptor signaling pathway / negative regulation of Wnt signaling pathway / positive regulation of activated T cell proliferation / negative regulation of translational frameshifting / TOR signaling / Protein hydroxylation / BH3 domain binding / SARS-CoV-1 modulates host translation machinery / regulation of adenylate cyclase-activating G protein-coupled receptor signaling pathway / iron-sulfur cluster binding / regulation of cell division / cellular response to ethanol / mTORC1-mediated signalling / Peptide chain elongation / Selenocysteine synthesis / ribosomal small subunit binding / Formation of a pool of free 40S subunits / positive regulation of intrinsic apoptotic signaling pathway by p53 class mediator / Eukaryotic Translation Termination / ubiquitin ligase inhibitor activity / positive regulation of GTPase activity / SRP-dependent cotranslational protein targeting to membrane / Response of EIF2AK4 (GCN2) to amino acid deficiency / protein serine/threonine kinase inhibitor activity / negative regulation of ubiquitin-dependent protein catabolic process / positive regulation of signal transduction by p53 class mediator / Viral mRNA Translation / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / GTP hydrolysis and joining of the 60S ribosomal subunit / L13a-mediated translational silencing of Ceruloplasmin expression / Major pathway of rRNA processing in the nucleolus and cytosol / phagocytic cup / regulation of translational fidelity / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / negative regulation of protein binding / Nuclear events stimulated by ALK signaling in cancer / positive regulation of intrinsic apoptotic signaling pathway 類似検索 - 分子機能 | |||||||||
| 生物種 | Homo sapiens (ヒト) | |||||||||
| 手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 6.3 Å | |||||||||
データ登録者 | Schaffitzel, C. / Schaffitzel, C. | |||||||||
| 資金援助 | ベルギー, 2件
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引用 | ジャーナル: Nucleic Acids Res / 年: 2018タイトル: Structure of a human cap-dependent 48S translation pre-initiation complex. 著者: Boris Eliseev / Lahari Yeramala / Alexander Leitner / Manikandan Karuppasamy / Etienne Raimondeau / Karine Huard / Elena Alkalaeva / Ruedi Aebersold / Christiane Schaffitzel / ![]() 要旨: Eukaryotic translation initiation is tightly regulated, requiring a set of conserved initiation factors (eIFs). Translation of a capped mRNA depends on the trimeric eIF4F complex and eIF4B to load ...Eukaryotic translation initiation is tightly regulated, requiring a set of conserved initiation factors (eIFs). Translation of a capped mRNA depends on the trimeric eIF4F complex and eIF4B to load the mRNA onto the 43S pre-initiation complex comprising 40S and initiation factors 1, 1A, 2, 3 and 5 as well as initiator-tRNA. Binding of the mRNA is followed by mRNA scanning in the 48S pre-initiation complex, until a start codon is recognised. Here, we use a reconstituted system to prepare human 48S complexes assembled on capped mRNA in the presence of eIF4B and eIF4F. The highly purified h-48S complexes are used for cross-linking/mass spectrometry, revealing the protein interaction network in this complex. We report the electron cryo-microscopy structure of the h-48S complex at 6.3 Å resolution. While the majority of eIF4B and eIF4F appear to be flexible with respect to the ribosome, additional density is detected at the entrance of the 40S mRNA channel which we attribute to the RNA-recognition motif of eIF4B. The eight core subunits of eIF3 are bound at the 40S solvent-exposed side, as well as the subunits eIF3d, eIF3b and eIF3i. elF2 and initiator-tRNA bound to the start codon are present at the 40S intersubunit side. This cryo-EM structure represents a molecular snap-shot revealing the h-48S complex following 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 | 分子: Molmil Jmol/JSmol |
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ダウンロードとリンク
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ダウンロード
| PDBx/mmCIF形式 | 6fec.cif.gz | 2.6 MB | 表示 | PDBx/mmCIF形式 |
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| PDB形式 | pdb6fec.ent.gz | 表示 | PDB形式 | |
| PDBx/mmJSON形式 | 6fec.json.gz | ツリー表示 | PDBx/mmJSON形式 | |
| その他 | その他のダウンロード |
-検証レポート
| 文書・要旨 | 6fec_validation.pdf.gz | 1.4 MB | 表示 | wwPDB検証レポート |
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| 文書・詳細版 | 6fec_full_validation.pdf.gz | 1.6 MB | 表示 | |
| XML形式データ | 6fec_validation.xml.gz | 258.2 KB | 表示 | |
| CIF形式データ | 6fec_validation.cif.gz | 434.7 KB | 表示 | |
| アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/fe/6fec ftp://data.pdbj.org/pub/pdb/validation_reports/fe/6fec | HTTPS FTP |
-関連構造データ
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リンク
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集合体
| 登録構造単位 | ![]()
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要素
-EUKARYOTIC TRANSLATION INITIATION FACTOR ... , 14種, 14分子 123456789PSduw
| #1: タンパク質 | 分子量: 164902.656 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) |
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| #2: タンパク質 | 分子量: 97923.547 Da / 分子数: 1 / Mutation: A577Y / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: hela |
| #3: タンパク質 | 分子量: 52281.633 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: hela / 参照: UniProt: P60228 |
| #4: タンパク質 | 分子量: 37846.730 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: Hela / 参照: ubiquitinyl hydrolase 1 |
| #5: タンパク質 | 分子量: 39952.281 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: hela |
| #6: タンパク質 | 分子量: 25129.709 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: hela / 参照: UniProt: Q9UBQ5 |
| #7: タンパク質 | 分子量: 66804.766 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: hela / 参照: UniProt: Q9Y262 |
| #8: タンパク質 | 分子量: 42555.832 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: hela / 参照: UniProt: Q7L2H7 |
| #9: タンパク質 | 分子量: 42203.555 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) |
| #19: タンパク質 | 分子量: 30633.297 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: hela |
| #22: タンパク質 | 分子量: 45862.441 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: hela / 参照: UniProt: P41091 |
| #32: タンパク質・ペプチド | 分子量: 2103.416 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) |
| #49: タンパク質 | 分子量: 72324.820 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: EIF4B / プラスミド: pFastBac_eIF4B発現宿主: ![]() 参照: UniProt: P23588 |
| #50: タンパク質 | 分子量: 124402.336 Da / 分子数: 1 / Mutation: D104E, Y124F / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: hela |
-RNA鎖 , 3種, 3分子 AFN
| #10: RNA鎖 | 分子量: 572789.812 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: hela |
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| #11: RNA鎖 | 分子量: 8238.953 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: beta globin derived / プラスミド: pET28a-MVHL-STOP2 / 発現宿主: ![]() |
| #18: RNA鎖 | 分子量: 24231.510 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: hela / 参照: GenBank: 174924 |
+40S ribosomal protein ... , 31種, 31分子 GHIJKLQRUVWXYZabcefghijklnoqrst
-タンパク質 , 2種, 2分子 mp
| #41: タンパク質 | 分子量: 34669.113 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: hela / 参照: UniProt: P63244 |
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| #44: タンパク質 | 分子量: 8358.903 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト) / 細胞株: hela / 参照: UniProt: P62979 |
-非ポリマー , 1種, 415分子 
| #51: 水 | ChemComp-HOH / |
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-詳細
| Has protein modification | Y |
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-実験情報
-実験
| 実験 | 手法: 電子顕微鏡法 |
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| EM実験 | 試料の集合状態: PARTICLE / 3次元再構成法: 単粒子再構成法 |
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試料調製
| 構成要素 |
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| 分子量 | 値: 2.0 MDa / 実験値: NO | ||||||||||||||||||||||||||||||
| 由来(天然) |
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| 緩衝液 | pH: 7.5 詳細: 20 mM Tris HCl, 50 mM KOAc, 2.5 mM MgCl2, 2 mM DTT, 0.25 mM spermidine 0.25 mM GMPPNP | ||||||||||||||||||||||||||||||
| 試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | ||||||||||||||||||||||||||||||
| 試料支持 | グリッドのタイプ: Quantifoil R2/2 | ||||||||||||||||||||||||||||||
| 急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 277 K |
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電子顕微鏡撮影
| 実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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| 顕微鏡 | モデル: FEI TITAN KRIOS |
| 電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: FLOOD BEAM |
| 電子レンズ | モード: BRIGHT FIELD / 倍率(公称値): 112000 X / 最大 デフォーカス(公称値): 4000 nm / 最小 デフォーカス(公称値): 1500 nm |
| 試料ホルダ | 凍結剤: NITROGEN 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
| 撮影 | 電子線照射量: 30 e/Å2 / 検出モード: INTEGRATING フィルム・検出器のモデル: FEI FALCON II (4k x 4k) |
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解析
| ソフトウェア | 名称: PHENIX / バージョン: 1.13_2998: / 分類: 精密化 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| EMソフトウェア |
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| CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 対称性 | 点対称性: C1 (非対称) | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 3次元再構成 | 解像度: 6.3 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 50604 / アルゴリズム: BACK PROJECTION / クラス平均像の数: 1 / 対称性のタイプ: POINT | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 原子モデル構築 | プロトコル: RIGID BODY FIT / 空間: REAL / Target criteria: Cross-correlation coefficient | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 精密化 | 解像度: 6.3→270 Å / Cor.coef. Fo:Fc: 0.961 / SU B: 135.967 / SU ML: 0.877 立体化学のターゲット値: MAXIMUM LIKELIHOOD WITH PHASES 詳細: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
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| 溶媒の処理 | イオンプローブ半径: 0.8 Å / 減衰半径: 0.8 Å / VDWプローブ半径: 1.2 Å / 溶媒モデル: MASK | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 原子変位パラメータ | Biso mean: 105.458 Å2
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| 精密化ステップ | サイクル: 1 / 合計: 32425 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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万見について




Homo sapiens (ヒト)
ベルギー, 2件
引用

UCSF Chimera









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