+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-3461 | |||||||||
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タイトル | Cryo-EM structure of a native ribosome-Ski2-Ski3-Ski8 complex from S. cerevisiae | |||||||||
マップデータ | Native ribosome-Ski2-Ski3-Ski8 complex from S. cerevisiae | |||||||||
試料 |
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機能・相同性 | 機能・相同性情報 positive regulation of cytoplasmic translational elongation through polyproline stretches / Hypusine synthesis from eIF5A-lysine / CAT tailing / translational frameshifting / Ski complex / mRNA decay by 3' to 5' exoribonuclease / protein-DNA complex assembly / positive regulation of translational termination / nuclear-transcribed mRNA catabolic process, 3'-5' exonucleolytic nonsense-mediated decay / : ...positive regulation of cytoplasmic translational elongation through polyproline stretches / Hypusine synthesis from eIF5A-lysine / CAT tailing / translational frameshifting / Ski complex / mRNA decay by 3' to 5' exoribonuclease / protein-DNA complex assembly / positive regulation of translational termination / nuclear-transcribed mRNA catabolic process, 3'-5' exonucleolytic nonsense-mediated decay / : / nuclear-transcribed mRNA catabolic process, non-stop decay / positive regulation of translational elongation / 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 / Protein methylation / RMTs methylate histone arginines / positive regulation of translational fidelity / reciprocal meiotic recombination / mTORC1-mediated signalling / ribosome-associated ubiquitin-dependent protein catabolic process / ヒドロキシル化 / GDP-dissociation inhibitor activity / : / pre-mRNA 5'-splice site binding / positive regulation of nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / Formation of the ternary complex, and subsequently, the 43S complex / Translation initiation complex formation / Ribosomal scanning and start codon recognition / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / preribosome, small subunit precursor / translational elongation / response to cycloheximide / nuclear chromosome / mRNA destabilization / Major pathway of rRNA processing in the nucleolus and cytosol / SRP-dependent cotranslational protein targeting to membrane / 90S preribosome / GTP hydrolysis and joining of the 60S ribosomal subunit / mRNA catabolic process / Formation of a pool of free 40S subunits / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / protein-RNA complex assembly / negative regulation of mRNA splicing, via spliceosome / ribosomal small subunit export from nucleus / positive regulation of translational initiation / preribosome, large subunit precursor / L13a-mediated translational silencing of Ceruloplasmin expression / translation regulator activity / ribosomal large subunit export from nucleus / G-protein alpha-subunit binding / 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) / regulation of translational fidelity / positive regulation of protein kinase activity / translation elongation factor activity / rescue of stalled ribosome / translational termination / maturation of SSU-rRNA / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of LSU-rRNA / ribosomal large subunit biogenesis / DNA-(apurinic or apyrimidinic site) endonuclease activity / cellular response to amino acid starvation / ribosome assembly / translation initiation factor activity / small-subunit processome / protein kinase C binding / maintenance of translational fidelity / オートファジー / modification-dependent protein catabolic process / ribosomal small subunit biogenesis / ribosomal large subunit assembly / small ribosomal subunit rRNA binding / protein tag activity / ribosomal small subunit assembly / rRNA processing / cytoplasmic stress granule / cytosolic small ribosomal subunit / large ribosomal subunit rRNA binding / ribosome binding / リボソーム生合成 / regulation of translation / small ribosomal subunit / cytoplasmic translation / 5S rRNA binding / cytosolic large ribosomal subunit / protein-containing complex assembly / defense response to virus / negative regulation of translation / RNA helicase activity / rRNA binding / protein ubiquitination / リボソーム / ヘリカーゼ / structural constituent of ribosome / positive regulation of protein phosphorylation / 翻訳 (生物学) 類似検索 - 分子機能 | |||||||||
生物種 | Saccharomyces cerevisiae (パン酵母) / Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (パン酵母) / Baker's yeast / Baker's yeast (パン酵母) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.8 Å | |||||||||
データ登録者 | Schmidt C / Kowalinski E / Shanmuganathan V / Defenouillere Q / Braunger K / Heuer A / Pech M / Namane A / Berninghausen O / Fromont-Racine M ...Schmidt C / Kowalinski E / Shanmuganathan V / Defenouillere Q / Braunger K / Heuer A / Pech M / Namane A / Berninghausen O / Fromont-Racine M / Jacquier A / Conti E / Becker T / Beckmann R | |||||||||
引用 | ジャーナル: Science / 年: 2016 タイトル: The cryo-EM structure of a ribosome-Ski2-Ski3-Ski8 helicase complex. 著者: Christian Schmidt / Eva Kowalinski / Vivekanandan Shanmuganathan / Quentin Defenouillère / Katharina Braunger / André Heuer / Markus Pech / Abdelkader Namane / Otto Berninghausen / ...著者: Christian Schmidt / Eva Kowalinski / Vivekanandan Shanmuganathan / Quentin Defenouillère / Katharina Braunger / André Heuer / Markus Pech / Abdelkader Namane / Otto Berninghausen / Micheline Fromont-Racine / Alain Jacquier / Elena Conti / Thomas Becker / Roland Beckmann / 要旨: Ski2-Ski3-Ski8 (Ski) is a helicase complex functioning with the RNA-degrading exosome to mediate the 3'-5' messenger RNA (mRNA) decay in turnover and quality-control pathways. We report that the Ski ...Ski2-Ski3-Ski8 (Ski) is a helicase complex functioning with the RNA-degrading exosome to mediate the 3'-5' messenger RNA (mRNA) decay in turnover and quality-control pathways. We report that the Ski complex directly associates with 80S ribosomes presenting a short mRNA 3' overhang. We determined the structure of an endogenous ribosome-Ski complex using cryo-electron microscopy (EM) with a local resolution of the Ski complex ranging from 4 angstroms (Å) in the core to about 10 Å for intrinsically flexible regions. Ribosome binding displaces the autoinhibitory domain of the Ski2 helicase, positioning it in an open conformation near the ribosomal mRNA entry tunnel. We observe that the mRNA 3' overhang is threaded directly from the small ribosomal subunit to the helicase channel of Ski2, primed for ongoing exosome-mediated 3'-5' degradation. | |||||||||
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_3461.map.gz | 381.9 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-3461-v30.xml emd-3461.xml | 99.1 KB 99.1 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_3461.png | 38.7 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-3461 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-3461 | 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_3461.map.gz / 形式: CCP4 / 大きさ: 421.9 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Native ribosome-Ski2-Ski3-Ski8 complex from S. cerevisiae | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.084 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 : Native ribosome-Ski2-Ski3-Ski8 complex from S. cerevisiae
+超分子 #1: Native ribosome-Ski2-Ski3-Ski8 complex from S. cerevisiae
+分子 #1: 18S ribosomal RNA
+分子 #35: mRNA
+分子 #36: A-site tRNA
+分子 #37: P-site tRNA
+分子 #83: 25S ribosomal RNA
+分子 #84: 5S ribosomal RNA
+分子 #85: 5.8S ribosomal RNA
+分子 #2: 40S ribosomal protein S3
+分子 #3: 40S ribosomal protein S5
+分子 #4: 40S ribosomal protein S10-A
+分子 #5: 40S ribosomal protein S12
+分子 #6: 40S ribosomal protein S15
+分子 #7: 40S ribosomal protein S16-A
+分子 #8: 40S ribosomal protein S17-B
+分子 #9: 40S ribosomal protein S18-A
+分子 #10: 40S ribosomal protein S19-A
+分子 #11: 40S ribosomal protein S20
+分子 #12: 40S ribosomal protein S25-A
+分子 #13: 40S ribosomal protein S28-B
+分子 #14: 40S ribosomal protein S29-A
+分子 #15: Ubiquitin-40S ribosomal protein S31
+分子 #16: Guanine nucleotide-binding protein subunit beta-like protein
+分子 #17: 40S ribosomal protein S0-A
+分子 #18: 40S ribosomal protein S1-A
+分子 #19: 40S ribosomal protein S2
+分子 #20: 40S ribosomal protein S4-A
+分子 #21: 40S ribosomal protein S6-A
+分子 #22: 40S ribosomal protein S7-A
+分子 #23: 40S ribosomal protein S8-A
+分子 #24: 40S ribosomal protein S9-A
+分子 #25: 40S ribosomal protein S11-A
+分子 #26: 40S ribosomal protein S13
+分子 #27: 40S ribosomal protein S14-A
+分子 #28: 40S ribosomal protein S21-A
+分子 #29: 40S ribosomal protein S22-A
+分子 #30: 40S ribosomal protein S23-A
+分子 #31: 40S ribosomal protein S24-A
+分子 #32: 40S ribosomal protein S26-A
+分子 #33: 40S ribosomal protein S27-A
+分子 #34: 40S ribosomal protein S30-A
+分子 #38: Antiviral helicase SKI2
+分子 #39: Superkiller protein 3
+分子 #40: Antiviral protein SKI8
+分子 #41: 60S ribosomal protein L8-A
+分子 #42: 60S ribosomal protein L23-A
+分子 #43: 60S ribosomal protein L36-A
+分子 #44: 60S ribosomal protein L9-A
+分子 #45: 60S ribosomal protein L24-A
+分子 #46: 60S ribosomal protein L37-A
+分子 #47: 60S ribosomal protein L11-A
+分子 #48: 60S ribosomal protein L25
+分子 #49: 60S ribosomal protein L38
+分子 #50: 60S ribosomal protein L13-A
+分子 #51: 60S ribosomal protein L26-A
+分子 #52: 60S ribosomal protein L39
+分子 #53: 60S ribosomal protein L14-A
+分子 #54: 60S ribosomal protein L27-A
+分子 #55: Ubiquitin-60S ribosomal protein L40
+分子 #56: 60S ribosomal protein L42-A
+分子 #57: 60S ribosomal protein L15-A
+分子 #58: 60S ribosomal protein L28
+分子 #59: 60S ribosomal protein L41-A
+分子 #60: 60S ribosomal protein L43-A
+分子 #61: 60S ribosomal protein L16-A
+分子 #62: 60S ribosomal protein L29
+分子 #63: 60S ribosomal protein L2-A
+分子 #64: 60S ribosomal protein L17-A
+分子 #65: 60S ribosomal protein L30
+分子 #66: uL1
+分子 #67: 60S ribosomal protein L3
+分子 #68: 60S ribosomal protein L18-A
+分子 #69: 60S ribosomal protein L31-A
+分子 #70: 60S ribosomal protein L10
+分子 #71: 60S ribosomal protein L4-A
+分子 #72: 60S ribosomal protein L19-A
+分子 #73: 60S ribosomal protein L32
+分子 #74: 60S ribosomal protein L20-A
+分子 #75: 60S ribosomal protein L5
+分子 #76: 60S ribosomal protein L21-A
+分子 #77: 60S ribosomal protein L33-A
+分子 #78: 60S ribosomal protein L22-A
+分子 #79: 60S ribosomal protein L6-A
+分子 #80: 60S ribosomal protein L34-A
+分子 #81: 60S ribosomal protein L7-A
+分子 #82: 60S ribosomal protein L35-A
+分子 #86: Eukaryotic translation initiation factor 5A-1
-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: 7.4 構成要素:
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グリッド | モデル: Quantifoil R3/3 / 材質: COPPER/PALLADIUM / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: CONTINUOUS / 支持フィルム - Film thickness: 2.0 nm / 前処理 - タイプ: GLOW DISCHARGE | |||||||||||||||||||||
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 278 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 / Cs: 2.7 mm |
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) デジタル化 - 画像ごとのフレーム数: 1-10 / 撮影したグリッド数: 2 / 平均電子線量: 2.8 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: OTHER / 詳細: Yeast 80S ribosome |
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 角度割当 | タイプ: ANGULAR RECONSTITUTION |
最終 再構成 | 想定した対称性 - 点群: C1 (非対称) / 解像度のタイプ: BY AUTHOR / 解像度: 3.8 Å / 解像度の算出法: FSC 0.143 CUT-OFF / ソフトウェア - 名称: FREALIGN (ver. 9.11) / 使用した粒子像数: 31503 |