+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-4242 | |||||||||
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タイトル | Human cap-dependent 48S pre-initiation complex | |||||||||
マップデータ | human 48S pre-initiation complex, reconstituted in the presence of capped mRNA, eIF4B and eIF4F, displaying eIF2 ternary complex, eIF3 and eIF4B | |||||||||
試料 |
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機能・相同性 | 機能・相同性情報 positive regulation of mRNA binding / viral translational termination-reinitiation / eukaryotic translation initiation factor 3 complex, eIF3e / methionyl-initiator methionine tRNA binding / eukaryotic translation initiation factor 3 complex, eIF3m / Response of EIF2AK1 (HRI) to heme deficiency / Activation of the mRNA upon binding of the cap-binding complex and eIFs, and subsequent binding to 43S / Recycling of eIF2:GDP / eukaryotic translation initiation factor 2 complex / PERK regulates gene expression ...positive regulation of mRNA binding / viral translational termination-reinitiation / eukaryotic translation initiation factor 3 complex, eIF3e / methionyl-initiator methionine tRNA binding / eukaryotic translation initiation factor 3 complex, eIF3m / Response of EIF2AK1 (HRI) to heme deficiency / Activation of the mRNA upon binding of the cap-binding complex and eIFs, and subsequent binding to 43S / Recycling of eIF2:GDP / eukaryotic translation initiation factor 2 complex / PERK regulates gene expression / eukaryotic translation initiation factor 3 complex / eukaryotic translation initiation factor 4F complex / Z-decay: degradation of maternal mRNAs by zygotically expressed factors / eukaryotic 43S preinitiation complex / protein-synthesizing GTPase / cytoplasmic translational initiation / translation factor activity, RNA binding / formation of translation preinitiation complex / formation of cytoplasmic translation initiation complex / Deadenylation of mRNA / positive regulation of cysteine-type endopeptidase activity involved in execution phase of apoptosis / negative regulation of endoplasmic reticulum unfolded protein response / oxidized pyrimidine DNA binding / response to TNF agonist / positive regulation of base-excision repair / protein tyrosine kinase inhibitor activity / eukaryotic 48S preinitiation complex / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage / positive regulation of gastrulation / IRE1-RACK1-PP2A complex / positive regulation of endodeoxyribonuclease activity / positive regulation of Golgi to plasma membrane protein transport / TNFR1-mediated ceramide production / M-decay: degradation of maternal mRNAs by maternally stored factors / laminin receptor activity / negative regulation of DNA repair / negative regulation of RNA splicing / negative regulation of intrinsic apoptotic signaling pathway in response to hydrogen peroxide / oxidized purine DNA binding / supercoiled DNA binding / neural crest cell differentiation / NF-kappaB complex / ubiquitin-like protein conjugating enzyme binding / regulation of establishment of cell polarity / negative regulation of phagocytosis / regulation of translational initiation / Formation of the ternary complex, and subsequently, the 43S complex / rRNA modification in the nucleus and cytosol / erythrocyte homeostasis / cytoplasmic side of rough endoplasmic reticulum membrane / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / 顔料 / protein kinase A binding / Ribosomal scanning and start codon recognition / negative regulation of ubiquitin protein ligase activity / ion channel inhibitor activity / Translation initiation complex formation / positive regulation of mitochondrial depolarization / negative regulation of Wnt signaling pathway / positive regulation of T cell receptor signaling pathway / positive regulation of activated T cell proliferation / regulation of cell division / ヒドロキシル化 / iron-sulfur cluster binding / BH3 domain binding / mTORC1-mediated signalling / SARS-CoV-1 modulates host translation machinery / Peptide chain elongation / positive regulation of intrinsic apoptotic signaling pathway by p53 class mediator / Selenocysteine synthesis / cysteine-type endopeptidase activator activity involved in apoptotic process / positive regulation of signal transduction by p53 class mediator / Formation of a pool of free 40S subunits / ubiquitin ligase inhibitor activity / positive regulation of cyclic-nucleotide phosphodiesterase activity / Eukaryotic Translation Termination / phagocytic cup / Response of EIF2AK4 (GCN2) to amino acid deficiency / SRP-dependent cotranslational protein targeting to membrane / Viral mRNA Translation / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / GTP hydrolysis and joining of the 60S ribosomal subunit / MTOR / L13a-mediated translational silencing of Ceruloplasmin expression / Major pathway of rRNA processing in the nucleolus and cytosol / regulation of translational fidelity / spindle assembly / negative regulation of phosphatidylinositol 3-kinase/protein kinase B signal transduction / ribosomal small subunit export from nucleus / Nuclear events stimulated by ALK signaling in cancer / laminin binding / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / translation regulator activity / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / positive regulation of cell cycle / negative regulation of smoothened signaling pathway / signaling adaptor activity / stress granule assembly / negative regulation of peptidyl-serine phosphorylation / positive regulation of intrinsic apoptotic signaling pathway 類似検索 - 分子機能 | |||||||||
生物種 | Homo sapiens (ヒト) / Human (ヒト) | |||||||||
手法 | 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 | |||||||||
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 | 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_4242.map.gz | 32.1 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-4242-v30.xml emd-4242.xml | 71.1 KB 71.1 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_4242.png | 56.4 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-4242 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-4242 | 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_4242.map.gz / 形式: CCP4 / 大きさ: 34.3 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | human 48S pre-initiation complex, reconstituted in the presence of capped mRNA, eIF4B and eIF4F, displaying eIF2 ternary complex, eIF3 and eIF4B | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 2.5 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
-試料の構成要素
+全体 : human cap-dependent 48S translation pre-initiation complex
+超分子 #1: human cap-dependent 48S translation pre-initiation complex
+超分子 #2: human cap-dependent 48S translation pre-initiation complex
+超分子 #3: mRNA
+超分子 #4: Eukaryotic translation initiation factor 4B
+分子 #1: Eukaryotic translation initiation factor 3 subunit A
+分子 #2: Eukaryotic translation initiation factor 3 subunit C
+分子 #3: Eukaryotic translation initiation factor 3 subunit E
+分子 #4: Eukaryotic translation initiation factor 3 subunit F
+分子 #5: Eukaryotic translation initiation factor 3 subunit H
+分子 #6: Eukaryotic translation initiation factor 3 subunit K
+分子 #7: Eukaryotic translation initiation factor 3 subunit L
+分子 #8: Eukaryotic translation initiation factor 3 subunit M
+分子 #9: EUKARYOTIC TRANSLATION INITIATION FACTOR 3 SUBUNIT D
+分子 #12: 40S ribosomal protein S11
+分子 #13: 40S ribosomal protein S16
+分子 #14: 40S ribosomal protein S4, X isoform
+分子 #15: 40S ribosomal protein S29
+分子 #16: 40S ribosomal protein S9
+分子 #17: 40S ribosomal protein S18
+分子 #19: Eukaryotic translation initiation factor 2 subunit 1
+分子 #20: 40S ribosomal protein S23
+分子 #21: 40S ribosomal protein S19
+分子 #22: Eukaryotic translation initiation factor 2 subunit 3
+分子 #23: 40S ribosomal protein S5
+分子 #24: 40S ribosomal protein S30
+分子 #25: 40S ribosomal protein S25
+分子 #26: 40S ribosomal protein S7
+分子 #27: 40S ribosomal protein S27
+分子 #28: 40S ribosomal protein S13
+分子 #29: 40S ribosomal protein S15a
+分子 #30: 40S ribosomal protein S21
+分子 #31: 40S ribosomal protein S2
+分子 #32: EUKARYOTIC TRANSLATION INITIATION FACTOR 2 BETA SUBUNIT (eIF2-Beta)
+分子 #33: 40S ribosomal protein S17
+分子 #34: 40S ribosomal protein SA
+分子 #35: 40S ribosomal protein S3
+分子 #36: 40S ribosomal protein S20
+分子 #37: 40S ribosomal protein S3a
+分子 #38: 40S ribosomal protein S14
+分子 #39: 40S ribosomal protein S26
+分子 #40: 40S ribosomal protein S28
+分子 #41: Receptor of activated protein C kinase 1
+分子 #42: 40S ribosomal protein S15
+分子 #43: 40S ribosomal protein S8
+分子 #44: Ubiquitin-40S ribosomal protein S27a
+分子 #45: 40S ribosomal protein S6
+分子 #46: 40S ribosomal protein S12
+分子 #47: 40S ribosomal protein S24
+分子 #48: 40S ribosomal protein S10
+分子 #49: Eukaryotic translation initiation factor 4B
+分子 #50: Eukaryotic translation initiation factor 3 subunit B
+分子 #10: 18S ribosomal RNA
+分子 #11: Messenger RNA (26-MER)
+分子 #18: Transfer RNA (75-MER)
+分子 #51: water
-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
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 |
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グリッド | モデル: Quantifoil R2/2 |
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 |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELDBright-field microscopy / 最大 デフォーカス(公称値): 4.0 µm / 最小 デフォーカス(公称値): 1.5 µm / 倍率(公称値): 112000 |
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 |
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 / 平均電子線量: 30.0 e/Å2 |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
-画像解析
CTF補正 | ソフトウェア - 名称: CTFFIND (ver. 4) |
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初期モデル | モデルのタイプ: EMDB MAP EMDB ID: |
初期 角度割当 | タイプ: PROJECTION MATCHING / ソフトウェア - 名称: RELION (ver. 1.4) |
最終 3次元分類 | ソフトウェア - 名称: RELION (ver. 1.4) |
最終 角度割当 | タイプ: PROJECTION MATCHING / ソフトウェア - 名称: RELION (ver. 1.4) |
最終 再構成 | 使用したクラス数: 1 / 想定した対称性 - 点群: C1 (非対称) / アルゴリズム: BACK PROJECTION / 解像度のタイプ: BY AUTHOR / 解像度: 6.3 Å / 解像度の算出法: FSC 0.143 CUT-OFF / ソフトウェア - 名称: RELION (ver. 1.4) / 使用した粒子像数: 50604 |
-原子モデル構築 1
精密化 | 空間: REAL / プロトコル: RIGID BODY FIT 当てはまり具合の基準: Cross-correlation coefficient |
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得られたモデル | PDB-6fec: |