+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-5942 | |||||||||
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タイトル | Cryo-EM Map of a yeast ribosome bound to the TSV IRES (Class II) | |||||||||
マップデータ | Reconstruction of yeast 80S ribosome bound with the Taura Syndrome Virus IRES | |||||||||
試料 |
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キーワード | Translation Initiation / Internal Ribosome Entry Site | |||||||||
機能・相同性 | 機能・相同性情報 maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, LSU-rRNA,5S) / negative regulation of glucose mediated signaling pathway / negative regulation of translational frameshifting / Negative regulators of DDX58/IFIH1 signaling / RMTs methylate histone arginines / positive regulation of translational fidelity / Protein methylation / mTORC1-mediated signalling / Protein hydroxylation / ribosome-associated ubiquitin-dependent protein catabolic process ...maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, LSU-rRNA,5S) / negative regulation of glucose mediated signaling pathway / negative regulation of translational frameshifting / Negative regulators of DDX58/IFIH1 signaling / RMTs methylate histone arginines / positive regulation of translational fidelity / Protein methylation / mTORC1-mediated signalling / Protein hydroxylation / ribosome-associated ubiquitin-dependent protein catabolic process / GDP-dissociation inhibitor activity / hexon binding / positive regulation of nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / nonfunctional rRNA decay / pre-mRNA 5'-splice site binding / preribosome, small subunit precursor / Formation of the ternary complex, and subsequently, the 43S complex / Translation initiation complex formation / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / Ribosomal scanning and start codon recognition / response to cycloheximide / Major pathway of rRNA processing in the nucleolus and cytosol / mRNA destabilization / SRP-dependent cotranslational protein targeting to membrane / GTP hydrolysis and joining of the 60S ribosomal subunit / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / Formation of a pool of free 40S subunits / negative regulation of mRNA splicing, via spliceosome / preribosome, large subunit precursor / regulation of amino acid metabolic process / L13a-mediated translational silencing of Ceruloplasmin expression / translational elongation / ribosomal large subunit export from nucleus / 90S preribosome / G-protein alpha-subunit binding / positive regulation of protein kinase activity / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / regulation of translational fidelity / Ub-specific processing proteases / protein-RNA complex assembly / ribosomal subunit export from nucleus / ribosomal small subunit export from nucleus / translation regulator activity / translational termination / 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) / DNA-(apurinic or apyrimidinic site) endonuclease activity / maturation of LSU-rRNA / cellular response to amino acid starvation / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / ribosome assembly / rescue of stalled ribosome / ribosomal large subunit biogenesis / 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 / macroautophagy / maintenance of translational fidelity / modification-dependent protein catabolic process / cytoplasmic stress granule / protein tag activity / rRNA processing / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / ribosome biogenesis / viral capsid / ribosome binding / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / negative regulation of translation / rRNA binding / ribosome / protein ubiquitination / structural constituent of ribosome / positive regulation of protein phosphorylation / G protein-coupled receptor signaling pathway / translation / negative regulation of gene expression / response to antibiotic / mRNA binding / ubiquitin protein ligase binding / host cell nucleus / nucleolus / mitochondrion / RNA binding / zinc ion binding / nucleoplasm / nucleus / metal ion binding / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | Saccharomyces cerevisiae (パン酵母) / Taura syndrome virus (ウイルス) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 6.1 Å | |||||||||
データ登録者 | Koh CS / Brilot AF / Grigorieff N / Korostelev AA | |||||||||
引用 | ジャーナル: Proc Natl Acad Sci U S A / 年: 2014 タイトル: Taura syndrome virus IRES initiates translation by binding its tRNA-mRNA-like structural element in the ribosomal decoding center. 著者: Cha San Koh / Axel F Brilot / Nikolaus Grigorieff / Andrei A Korostelev / 要旨: In cap-dependent translation initiation, the open reading frame (ORF) of mRNA is established by the placement of the AUG start codon and initiator tRNA in the ribosomal peptidyl (P) site. Internal ...In cap-dependent translation initiation, the open reading frame (ORF) of mRNA is established by the placement of the AUG start codon and initiator tRNA in the ribosomal peptidyl (P) site. Internal ribosome entry sites (IRESs) promote translation of mRNAs in a cap-independent manner. We report two structures of the ribosome-bound Taura syndrome virus (TSV) IRES belonging to the family of Dicistroviridae intergenic IRESs. Intersubunit rotational states differ in these structures, suggesting that ribosome dynamics play a role in IRES translocation. Pseudoknot I of the IRES occupies the ribosomal decoding center at the aminoacyl (A) site in a manner resembling that of the tRNA anticodon-mRNA codon. The structures reveal that the TSV IRES initiates translation by a previously unseen mechanism, which is conceptually distinct from initiator tRNA-dependent mechanisms. Specifically, the ORF of the IRES-driven mRNA is established by the placement of the preceding tRNA-mRNA-like structure in the A site, whereas the 40S P site remains unoccupied during this initial step. | |||||||||
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_5942.map.gz | 253.9 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-5942-v30.xml emd-5942.xml | 14.2 KB 14.2 KB | 表示 表示 | EMDBヘッダ |
画像 | 400_5942.gif 80_5942.gif | 70.5 KB 4.6 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-5942 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-5942 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_5942_validation.pdf.gz | 346.6 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_5942_full_validation.pdf.gz | 346.2 KB | 表示 | |
XML形式データ | emd_5942_validation.xml.gz | 7.3 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-5942 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-5942 | HTTPS FTP |
-関連構造データ
関連構造データ | 3j6xMC 5943C 3j6yC C: 同じ文献を引用 (文献) M: このマップから作成された原子モデル |
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類似構造データ | |
電子顕微鏡画像生データ | EMPIAR-10015 (タイトル: Yeast 80S Ribosome-Taura Syndrome Virus IRES complex, Frealign Input Particle Stack Data size: 273.6 Data #1: Frealign input particle stack [picked particles - multiframe - processed]) |
-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_5942.map.gz / 形式: CCP4 / 大きさ: 276 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Reconstruction of yeast 80S ribosome bound with the Taura Syndrome Virus IRES | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.0595 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 : Saccharomyces cerevisiae 80S ribosome bound with TSV IRES
全体 | 名称: Saccharomyces cerevisiae 80S ribosome bound with TSV IRES |
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要素 |
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-超分子 #1000: Saccharomyces cerevisiae 80S ribosome bound with TSV IRES
超分子 | 名称: Saccharomyces cerevisiae 80S ribosome bound with TSV IRES タイプ: sample / ID: 1000 / Number unique components: 2 |
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分子量 | 実験値: 3.5 MDa / 理論値: 3.5 MDa |
-超分子 #1: 80S ribosome
超分子 | 名称: 80S ribosome / タイプ: complex / ID: 1 / 組換発現: No / データベース: NCBI / Ribosome-details: ribosome-eukaryote: ALL |
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由来(天然) | 生物種: Saccharomyces cerevisiae (パン酵母) / 株: W303 / 別称: Yeast |
分子量 | 実験値: 3.4 MDa / 理論値: 3.4 MDa |
-分子 #1: Internal Ribosome Entry Site
分子 | 名称: Internal Ribosome Entry Site / タイプ: rna / ID: 1 / Name.synonym: IRES / 分類: OTHER / Structure: DOUBLE HELIX / Synthetic?: Yes |
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由来(天然) | 生物種: Taura syndrome virus (ウイルス) |
分子量 | 実験値: 80 KDa / 理論値: 80 KDa |
配列 | 文字列: UAGCACCACC CGAUCGUAAA CUCCAUGUAU UGGUUACCCA UCUGCAUCGA AAACUCUCCG AACACUAGGU GCAGUAAGGC UUUCAUGGAG UGGUUUGCUA UUUAGCGUAC GUGUACCAUA GGCAGCCCCA AAAACACGUG UGAGGAGAAA GUCCCAGUCA CUUUGGGCAA ...文字列: UAGCACCACC CGAUCGUAAA CUCCAUGUAU UGGUUACCCA UCUGCAUCGA AAACUCUCCG AACACUAGGU GCAGUAAGGC UUUCAUGGAG UGGUUUGCUA UUUAGCGUAC GUGUACCAUA GGCAGCCCCA AAAACACGUG UGAGGAGAAA GUCCCAGUCA CUUUGGGCAA AGUAGACAGC CGCGCUUGCG UGGUGGGACU UAAUUAAUGC CUGCUAACCC AGUUGAAAUU GAUAAUUUUG AUACAACAAC |
-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 | 1.044 mg/mL |
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緩衝液 | pH: 7.5 詳細: 45 mM HEPES/KOH, 10 mM MgCl2, 100 mM KCl, 2.5 mM spermine, 2 mM BME, 0.5 U/ul RNasin |
グリッド | 詳細: C-flat 1.2-1.3 400C |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 95 % / 装置: FEI VITROBOT MARK II / 手法: Fresh glow discharge, 7 second blot |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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アライメント法 | Legacy - 非点収差: Corrected using FEI software bundled with Titan Krios/Cs Corrector. |
日付 | 2012年12月26日 |
撮影 | カテゴリ: CCD フィルム・検出器のモデル: FEI FALCON I (4k x 4k) デジタル化 - サンプリング間隔: 14.0 µm / 実像数: 11498 / 平均電子線量: 30 e/Å2 / ビット/ピクセル: 32 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 倍率(補正後): 132138 / 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): 6.53 µm / 最小 デフォーカス(公称値): 1.15 µm / 倍率(公称値): 133333 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
-画像解析
CTF補正 | 詳細: CTFFIND3, FREALIGN per micrograph |
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最終 再構成 | アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 6.1 Å / 解像度の算出法: OTHER ソフトウェア - 名称: EMAN2, IMAGIC, FREALIGN, RSAMPLE, CTFFIND3 使用した粒子像数: 52444 |
-原子モデル構築 1
初期モデル | PDB ID: 3u5b |
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ソフトウェア | 名称: Chimera, CNS |
詳細 | 1. TSV IRES was modeled with iFoldRNA and ModeRNA programs and then fitted into the cryo-EM map in Chimera. 2. The 80S-IRES complex was refined against the map, using stereochemically restrained real-space refinement in RSRef. |
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 |
得られたモデル | PDB-3j6x: |
-原子モデル構築 2
初期モデル | PDB ID: 3u5c |
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ソフトウェア | 名称: Chimera, CNS |
詳細 | 1. TSV IRES was modeled with iFoldRNA and ModeRNA programs and then fitted into the cryo-EM map in Chimera. 2. The 80S-IRES complex was refined against the map, using stereochemically restrained real-space refinement in RSRef. |
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 |
得られたモデル | PDB-3j6x: |
-原子モデル構築 3
初期モデル | PDB ID: 3u5d |
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ソフトウェア | 名称: Chimera, CNS |
詳細 | 1. TSV IRES was modeled with iFoldRNA and ModeRNA programs and then fitted into the cryo-EM map in Chimera. 2. The 80S-IRES complex was refined against the map, using stereochemically restrained real-space refinement in RSRef. |
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 |
得られたモデル | PDB-3j6x: |
-原子モデル構築 4
初期モデル | PDB ID: 3u5e |
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ソフトウェア | 名称: Chimera, CNS |
詳細 | 1. TSV IRES was modeled with iFoldRNA and ModeRNA programs and then fitted into the cryo-EM map in Chimera. 2. The 80S-IRES complex was refined against the map, using stereochemically restrained real-space refinement in RSRef. |
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 |
得られたモデル | PDB-3j6x: |