+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 4v7e | |||||||||
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タイトル | Model of the small subunit RNA based on a 5.5 A cryo-EM map of Triticum aestivum translating 80S ribosome | |||||||||
要素 |
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キーワード | RIBOSOME / eukaryotic ribosome / homology modeling / de novo modeling / ribosomal RNA / rRNA / RNA expansion segments | |||||||||
機能・相同性 | 機能・相同性情報 protein kinase activator activity / translational elongation / ribonucleoprotein complex binding / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / rRNA processing / ribosomal small subunit biogenesis / heterotrimeric G-protein complex / signaling receptor complex adaptor activity ...protein kinase activator activity / translational elongation / ribonucleoprotein complex binding / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / rRNA processing / ribosomal small subunit biogenesis / heterotrimeric G-protein complex / signaling receptor complex adaptor activity / ribosome biogenesis / ribosomal small subunit assembly / small ribosomal subunit / large ribosomal subunit rRNA binding / cytosolic small ribosomal subunit / cytoplasmic translation / cytosolic large ribosomal subunit / negative regulation of translation / ribosome / structural constituent of ribosome / G protein-coupled receptor signaling pathway / ribonucleoprotein complex / translation / mRNA binding / RNA binding / zinc ion binding / nucleus / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | Triticum aestivum (コムギ) | |||||||||
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: 5.5 Å | |||||||||
データ登録者 | Barrio-Garcia, C. / Armache, J.-P. / Jarasch, A. / Anger, A.M. / Villa, E. / Becker, T. / Bhushan, S. / Jossinet, F. / Habeck, M. / Dindar, G. ...Barrio-Garcia, C. / Armache, J.-P. / Jarasch, A. / Anger, A.M. / Villa, E. / Becker, T. / Bhushan, S. / Jossinet, F. / Habeck, M. / Dindar, G. / Franckenberg, S. / Marquez, V. / Mielke, T. / Thomm, M. / Berninghausen, O. / Beatrix, B. / Soeding, J. / Westhof, E. / Wilson, D.N. / Beckmann, R. | |||||||||
引用 | ジャーナル: Nature / 年: 2014 タイトル: Structures of the Sec61 complex engaged in nascent peptide translocation or membrane insertion. 著者: Marko Gogala / Thomas Becker / Birgitta Beatrix / Jean-Paul Armache / Clara Barrio-Garcia / Otto Berninghausen / Roland Beckmann / 要旨: The biogenesis of secretory as well as transmembrane proteins requires the activity of the universally conserved protein-conducting channel (PCC), the Sec61 complex (SecY complex in bacteria). In ...The biogenesis of secretory as well as transmembrane proteins requires the activity of the universally conserved protein-conducting channel (PCC), the Sec61 complex (SecY complex in bacteria). In eukaryotic cells the PCC is located in the membrane of the endoplasmic reticulum where it can bind to translating ribosomes for co-translational protein transport. The Sec complex consists of three subunits (Sec61α, β and γ) and provides an aqueous environment for the translocation of hydrophilic peptides as well as a lateral opening in the Sec61α subunit that has been proposed to act as a gate for the membrane partitioning of hydrophobic domains. A plug helix and a so-called pore ring are believed to seal the PCC against ion flow and are proposed to rearrange for accommodation of translocating peptides. Several crystal and cryo-electron microscopy structures revealed different conformations of closed and partially open Sec61 and SecY complexes. However, in none of these samples has the translocation state been unambiguously defined biochemically. Here we present cryo-electron microscopy structures of ribosome-bound Sec61 complexes engaged in translocation or membrane insertion of nascent peptides. Our data show that a hydrophilic peptide can translocate through the Sec complex with an essentially closed lateral gate and an only slightly rearranged central channel. Membrane insertion of a hydrophobic domain seems to occur with the Sec complex opening the proposed lateral gate while rearranging the plug to maintain an ion permeability barrier. Taken together, we provide a structural model for the basic activities of the Sec61 complex as a protein-conducting channel. #1: ジャーナル: Proc Natl Acad Sci U S A / 年: 2010 タイトル: Cryo-EM structure and rRNA model of a translating eukaryotic 80S ribosome at 5.5-A resolution. 著者: Jean-Paul Armache / Alexander Jarasch / Andreas M Anger / Elizabeth Villa / Thomas Becker / Shashi Bhushan / Fabrice Jossinet / Michael Habeck / Gülcin Dindar / Sibylle Franckenberg / Viter ...著者: Jean-Paul Armache / Alexander Jarasch / Andreas M Anger / Elizabeth Villa / Thomas Becker / Shashi Bhushan / Fabrice Jossinet / Michael Habeck / Gülcin Dindar / Sibylle Franckenberg / Viter Marquez / Thorsten Mielke / Michael Thomm / Otto Berninghausen / Birgitta Beatrix / Johannes Söding / Eric Westhof / Daniel N Wilson / Roland Beckmann / 要旨: Protein biosynthesis, the translation of the genetic code into polypeptides, occurs on ribonucleoprotein particles called ribosomes. Although X-ray structures of bacterial ribosomes are available, ...Protein biosynthesis, the translation of the genetic code into polypeptides, occurs on ribonucleoprotein particles called ribosomes. Although X-ray structures of bacterial ribosomes are available, high-resolution structures of eukaryotic 80S ribosomes are lacking. Using cryoelectron microscopy and single-particle reconstruction, we have determined the structure of a translating plant (Triticum aestivum) 80S ribosome at 5.5-Å resolution. This map, together with a 6.1-Å map of a Saccharomyces cerevisiae 80S ribosome, has enabled us to model ∼98% of the rRNA. Accurate assignment of the rRNA expansion segments (ES) and variable regions has revealed unique ES-ES and r-protein-ES interactions, providing insight into the structure and evolution of the eukaryotic ribosome. #2: ジャーナル: Proc Natl Acad Sci U S A / 年: 2010 タイトル: Localization of eukaryote-specific ribosomal proteins in a 5.5-Å cryo-EM map of the 80S eukaryotic ribosome. 著者: Jean-Paul Armache / Alexander Jarasch / Andreas M Anger / Elizabeth Villa / Thomas Becker / Shashi Bhushan / Fabrice Jossinet / Michael Habeck / Gülcin Dindar / Sibylle Franckenberg / Viter ...著者: Jean-Paul Armache / Alexander Jarasch / Andreas M Anger / Elizabeth Villa / Thomas Becker / Shashi Bhushan / Fabrice Jossinet / Michael Habeck / Gülcin Dindar / Sibylle Franckenberg / Viter Marquez / Thorsten Mielke / Michael Thomm / Otto Berninghausen / Birgitta Beatrix / Johannes Söding / Eric Westhof / Daniel N Wilson / Roland Beckmann / 要旨: Protein synthesis in all living organisms occurs on ribonucleoprotein particles, called ribosomes. Despite the universality of this process, eukaryotic ribosomes are significantly larger in size than ...Protein synthesis in all living organisms occurs on ribonucleoprotein particles, called ribosomes. Despite the universality of this process, eukaryotic ribosomes are significantly larger in size than their bacterial counterparts due in part to the presence of 80 r proteins rather than 54 in bacteria. Using cryoelectron microscopy reconstructions of a translating plant (Triticum aestivum) 80S ribosome at 5.5-Å resolution, together with a 6.1-Å map of a translating Saccharomyces cerevisiae 80S ribosome, we have localized and modeled 74/80 (92.5%) of the ribosomal proteins, encompassing 12 archaeal/eukaryote-specific small subunit proteins as well as the complete complement of the ribosomal proteins of the eukaryotic large subunit. Near-complete atomic models of the 80S ribosome provide insights into the structure, function, and evolution of the eukaryotic translational apparatus. | |||||||||
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: MolmilJmol/JSmol |
-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 4v7e.cif.gz | 4.7 MB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb4v7e.ent.gz | 表示 | PDB形式 | |
PDBx/mmJSON形式 | 4v7e.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 | 4v7e_validation.pdf.gz | 1.8 MB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 4v7e_full_validation.pdf.gz | 3.4 MB | 表示 | |
XML形式データ | 4v7e_validation.xml.gz | 474.5 KB | 表示 | |
CIF形式データ | 4v7e_validation.cif.gz | 769.6 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/v7/4v7e ftp://data.pdbj.org/pub/pdb/validation_reports/v7/4v7e | HTTPS FTP |
-関連構造データ
-リンク
-集合体
登録構造単位 |
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-要素
-RNA鎖 , 6種, 6分子 AdAeAfAaAcAb
#1: RNA鎖 | 分子量: 583613.562 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Triticum aestivum (コムギ) / 参照: GenBank: AY049040.1 |
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#2: RNA鎖 | 分子量: 24135.262 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Triticum aestivum (コムギ) / 参照: GenBank: M25599.1 |
#3: RNA鎖 | 分子量: 3498.178 Da / 分子数: 1 / 由来タイプ: 天然 / 詳細: messenger RNA / 由来: (天然) Triticum aestivum (コムギ) |
#84: RNA鎖 | 分子量: 1097289.250 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Triticum aestivum (コムギ) |
#85: RNA鎖 | 分子量: 51535.613 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Triticum aestivum (コムギ) |
#86: RNA鎖 | 分子量: 38716.957 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Triticum aestivum (コムギ) / 参照: GenBank: X06094.1 |
+40S ribosomal protein ... , 32種, 32分子 BYBIBKBMBfBXBDBEBFBQBUBOBSBNBLBTBPBZBcBWBdBbBeBABRBBBVBaBJBCBGBH
-タンパク質 , 3種, 4分子 BgCsCtCq
#10: タンパク質 | 分子量: 41794.809 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Triticum aestivum (コムギ) / 参照: UniProt: Q8LNY6 | ||
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#67: タンパク質 | 分子量: 11596.995 Da / 分子数: 2 / 由来タイプ: 天然 / 由来: (天然) Triticum aestivum (コムギ) #70: タンパク質 | | 分子量: 34418.574 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Triticum aestivum (コムギ) |
+60S ribosomal protein ... , 45種, 46分子 CGCTCZCzCACJCHCVCNCaCQCDCRCPCXCWCYCrCcCdCeCjClCoCMCSCUCiCKCu...
-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 |
-試料調製
構成要素 |
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分子量 | 値: 4.2 MDa / 実験値: YES | ||||||||||||
緩衝液 | 名称: 20 mM HEPES/KOH, pH 7.5, 100 mM KOAc, 10 mM Mg(OAc)2, 0.01 mg/mL cycloheximide, 1 mM DTT, 0.01% Nikkol pH: 7.5 詳細: 20 mM HEPES/KOH, pH 7.5, 100 mM KOAc, 10 mM Mg(OAc)2, 0.01 mg/mL cycloheximide, 1 mM DTT, 0.01% Nikkol | ||||||||||||
試料 | 濃度: 0.02 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | ||||||||||||
試料支持 | 詳細: Quantifoil Grid with 2 nm carbon on top | ||||||||||||
急速凍結 | 装置: FEI VITROBOT MARK I / 凍結剤: ETHANE / 湿度: 100 % 詳細: Blot for 10 seconds (using 2 layers of filter paper) before plunging into liquid ethane (FEI VITROBOT). 手法: Blot for 10 seconds before plunging, use 2 layers of filter paper |
-電子顕微鏡撮影
実験機器 | モデル: Tecnai F30 / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TECNAI F30 |
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): 39000 X / 倍率(補正後): 38900 X / 最大 デフォーカス(公称値): 4500 nm / 最小 デフォーカス(公称値): 1000 nm / Cs: 2.26 mm |
試料ホルダ | 試料ホルダーモデル: OTHER / 資料ホルダタイプ: FEI Polara Cartridge System |
撮影 | 電子線照射量: 25 e/Å2 / フィルム・検出器のモデル: KODAK SO-163 FILM 詳細: Scanned at 5334 dpi on a Heidelberg Primescan Drum Scanner |
画像スキャン | デジタル画像の数: 1374 |
放射 | プロトコル: SINGLE WAVELENGTH / 単色(M)・ラウエ(L): M / 散乱光タイプ: x-ray |
放射波長 | 相対比: 1 |
-解析
EMソフトウェア | 名称: SPIDER / カテゴリ: 3次元再構成 | ||||||||||||
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CTF補正 | 詳細: Wiener Filter on 3D volumes (SPIDER) | ||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||
3次元再構成 | 手法: Projection Matching / 解像度: 5.5 Å / 解像度の算出法: FSC 0.5 CUT-OFF / 粒子像の数: 2108230 / ピクセルサイズ(公称値): 1.24 Å / ピクセルサイズ(実測値): 1.24 Å / 対称性のタイプ: POINT | ||||||||||||
精密化ステップ | サイクル: LAST
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