+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 5apo | ||||||
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タイトル | Structure of the yeast 60S ribosomal subunit in complex with Arx1, Alb1 and C-terminally tagged Rei1 | ||||||
要素 |
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キーワード | RIBOSOME / EUKARYOTIC RIBOSOME BIOGENESIS / RIBOSOME MATURATION / RIBOSOME BIOGENESIS FACTOR / 60S RIBOSOMAL SUBUNIT / REI1 / ARX1 / ALB1 / CRYO-EM / RIBOSOMAL POLYPEPTIDE EXIT TUNNEL PROOFREADING | ||||||
機能・相同性 | 機能・相同性情報 加水分解酵素 / hexon binding / pre-mRNA 5'-splice site binding / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / response to cycloheximide / 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 ...加水分解酵素 / hexon binding / pre-mRNA 5'-splice site binding / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / response to cycloheximide / 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 / L13a-mediated translational silencing of Ceruloplasmin expression / translational elongation / ribosomal large subunit export from nucleus / 90S preribosome / regulation of translational fidelity / protein-RNA complex assembly / translational termination / maturation of LSU-rRNA / Neutrophil degranulation / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / ribosomal large subunit biogenesis / translational initiation / macroautophagy / maintenance of translational fidelity / modification-dependent protein catabolic process / protein tag activity / rRNA processing / metallopeptidase activity / ribosome biogenesis / viral capsid / 5S rRNA binding / large ribosomal subunit rRNA binding / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / negative regulation of translation / rRNA binding / ribosome / protein ubiquitination / structural constituent of ribosome / translation / response to antibiotic / mRNA binding / ubiquitin protein ligase binding / host cell nucleus / nucleolus / proteolysis / RNA binding / nucleoplasm / nucleus / metal ion binding / cytosol / cytoplasm 類似検索 - 分子機能 | ||||||
生物種 | Saccharomyces cerevisiae S288c (パン酵母) Saccharomyces cerevisiae S288C (パン酵母) | ||||||
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: 3.41 Å | ||||||
データ登録者 | Greber, B.J. / Gerhardy, S. / Leitner, A. / Leibundgut, M. / Salem, M. / Boehringer, D. / Leulliot, N. / Aebersold, R. / Panse, V.G. / Ban, N. | ||||||
引用 | ジャーナル: Cell / 年: 2016 タイトル: Insertion of the Biogenesis Factor Rei1 Probes the Ribosomal Tunnel during 60S Maturation. 著者: Basil Johannes Greber / Stefan Gerhardy / Alexander Leitner / Marc Leibundgut / Michèle Salem / Daniel Boehringer / Nicolas Leulliot / Ruedi Aebersold / Vikram Govind Panse / Nenad Ban / 要旨: Eukaryotic ribosome biogenesis depends on several hundred assembly factors to produce functional 40S and 60S ribosomal subunits. The final phase of 60S subunit biogenesis is cytoplasmic maturation, ...Eukaryotic ribosome biogenesis depends on several hundred assembly factors to produce functional 40S and 60S ribosomal subunits. The final phase of 60S subunit biogenesis is cytoplasmic maturation, which includes the proofreading of functional centers of the 60S subunit and the release of several ribosome biogenesis factors. We report the cryo-electron microscopy (cryo-EM) structure of the yeast 60S subunit in complex with the biogenesis factors Rei1, Arx1, and Alb1 at 3.4 Å resolution. In addition to the network of interactions formed by Alb1, the structure reveals a mechanism for ensuring the integrity of the ribosomal polypeptide exit tunnel. Arx1 probes the entire set of inner-ring proteins surrounding the tunnel exit, and the C terminus of Rei1 is deeply inserted into the ribosomal tunnel, where it forms specific contacts along almost its entire length. We provide genetic and biochemical evidence that failure to insert the C terminus of Rei1 precludes subsequent steps of 60S maturation. | ||||||
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 | 5apo.cif.gz | 3.2 MB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb5apo.ent.gz | 表示 | PDB形式 | |
PDBx/mmJSON形式 | 5apo.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 | 5apo_validation.pdf.gz | 1.2 MB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 5apo_full_validation.pdf.gz | 1.3 MB | 表示 | |
XML形式データ | 5apo_validation.xml.gz | 207.1 KB | 表示 | |
CIF形式データ | 5apo_validation.cif.gz | 366 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/ap/5apo ftp://data.pdbj.org/pub/pdb/validation_reports/ap/5apo | 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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1 |
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-要素
-RNA鎖 , 3種, 3分子 578
#1: RNA鎖 | 分子量: 1094331.625 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) Saccharomyces cerevisiae S288c (パン酵母) 参照: GenBank: 329138943 |
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#2: RNA鎖 | 分子量: 38951.105 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) Saccharomyces cerevisiae S288c (パン酵母) 参照: GenBank: 329138943 |
#3: RNA鎖 | 分子量: 50682.922 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) Saccharomyces cerevisiae S288c (パン酵母) 参照: GenBank: 329138943 |
+60S ribosomal protein ... , 39種, 39分子 ABCDEFGHIJLMNOPQRSTUVWXYZabcde...
-タンパク質 , 5種, 5分子 mqxyz
#41: タンパク質 | 分子量: 14583.077 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) Saccharomyces cerevisiae S288c (パン酵母) 参照: UniProt: P0CH08 |
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#44: タンパク質 | 分子量: 33749.121 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) Saccharomyces cerevisiae S288c (パン酵母) 参照: UniProt: P05317 |
#45: タンパク質 | 分子量: 67835.188 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) Saccharomyces cerevisiae S288c (パン酵母) 発現宿主: Escherichia coli BL21(DE3) (大腸菌) / Variant (発現宿主): Rosetta 2 / 参照: UniProt: Q03862, 加水分解酵素 |
#46: タンパク質 | 分子量: 46541.117 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) Saccharomyces cerevisiae S288c (パン酵母) 発現宿主: Escherichia coli BL21(DE3) (大腸菌) / Variant (発現宿主): Rosetta 2 |
#47: タンパク質 | 分子量: 8102.979 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) Saccharomyces cerevisiae S288c (パン酵母) 発現宿主: Escherichia coli BL21(DE3) (大腸菌) / Variant (発現宿主): Rosetta 2 |
-非ポリマー , 2種, 286分子
#48: 化合物 | ChemComp-MG / #49: 化合物 | ChemComp-ZN / |
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-詳細
Has protein modification | N |
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配列の詳細 | The correct register of the chain z remains unknown for some parts of the model. Hence they are ...The correct register of the chain z remains unknown for some parts of the model. Hence they are built in as UNK. However, the sample sequence for chain z corresponds to UniProt P47019.1 |
-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 | 名称: YEAST 60S ARX1 ALB1 REI1- 6XHIS / タイプ: RIBOSOME 詳細: QUANTIFOIL HOLEY CARBON GRIDS WERE COATED WITH A THIN CARBON FILM |
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緩衝液 | 名称: 20MM HEPES-KOH, 100mM NACL, 5mM MGCL2, 5mM BETA-MERCAPTOETHANOL pH: 8 詳細: 20MM HEPES-KOH, 100mM NACL, 5mM MGCL2, 5mM BETA-MERCAPTOETHANOL |
試料 | 濃度: 0.2 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: HOLEY CARBON |
急速凍結 | 装置: HOMEMADE PLUNGER / 凍結剤: ETHANE-PROPANE / 詳細: PLUNGE-FROZEN |
-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 / 日付: 2015年4月7日 / 詳細: Collected in movie mode in 2 sessions |
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): 59000 X / 倍率(補正後): 100720 X / 最大 デフォーカス(公称値): 3000 nm / 最小 デフォーカス(公称値): 800 nm / Cs: 2.7 mm |
試料ホルダ | 温度: 80 K |
撮影 | 電子線照射量: 20 e/Å2 フィルム・検出器のモデル: FEI FALCON II (4k x 4k) |
画像スキャン | デジタル画像の数: 3654 |
-解析
EMソフトウェア |
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CTF補正 | 詳細: PER DETECTOR FRAME | ||||||||||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||||||||||
3次元再構成 | 手法: MAXIMUM LIKELIHOOD BASED REFINEMENT IMPLEMENTED IN RELION 解像度: 3.41 Å / 粒子像の数: 134701 / ピクセルサイズ(実測値): 1.39 Å 詳細: FOR VISUALIZATION PURPOSES THE FINAL MAP WAS FILTERED AND AMPLITUDE CORRECTED IN RELION. SUBMISSION BASED ON EXPERIMENTAL DATA FROM EMDB EMD-3151. THE COORDINATES WERE REFINED IN RECIPROCAL ...詳細: FOR VISUALIZATION PURPOSES THE FINAL MAP WAS FILTERED AND AMPLITUDE CORRECTED IN RELION. SUBMISSION BASED ON EXPERIMENTAL DATA FROM EMDB EMD-3151. THE COORDINATES WERE REFINED IN RECIPROCAL SPACE USING PHENIX.REFINE AGAINST THE MLHL TARGET. FOR THIS, THE CRYO-EM MAP WAS CONVERTED TO RECIPROCAL SPACE STRUCTURE FACTORS. 対称性のタイプ: POINT | ||||||||||||||||||||
原子モデル構築 | 空間: REAL | ||||||||||||||||||||
精密化 | 最高解像度: 3.41 Å |