+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 4d61 | |||||||||
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タイトル | Cryo-EM structures of ribosomal 80S complexes with termination factors and cricket paralysis virus IRES reveal the IRES in the translocated state | |||||||||
要素 |
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キーワード | RIBOSOME / CRPV IRES / TERMINATION / RELEASE FACTORS | |||||||||
機能・相同性 | 機能・相同性情報 translation termination factor activity / cytoplasmic translational termination / translation release factor complex / translation release factor activity / regulation of translational termination / ribosomal subunit / translation release factor activity, codon specific / protein methylation / sequence-specific mRNA binding / aminoacyl-tRNA hydrolase activity ...translation termination factor activity / cytoplasmic translational termination / translation release factor complex / translation release factor activity / regulation of translational termination / ribosomal subunit / translation release factor activity, codon specific / protein methylation / sequence-specific mRNA binding / aminoacyl-tRNA hydrolase activity / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / laminin receptor activity / mammalian oogenesis stage / activation-induced cell death of T cells / Protein hydroxylation / positive regulation of signal transduction by p53 class mediator / ubiquitin ligase inhibitor activity / Eukaryotic Translation Termination / phagocytic cup / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / 90S preribosome / TOR signaling / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / T cell proliferation involved in immune response / erythrocyte development / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / ribosomal small subunit export from nucleus / translation regulator activity / translational termination / laminin binding / rough endoplasmic reticulum / 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) / gastrulation / MDM2/MDM4 family protein binding / cytosolic ribosome / class I DNA-(apurinic or apyrimidinic site) endonuclease activity / DNA-(apurinic or apyrimidinic site) lyase / rescue of stalled ribosome / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / cellular response to leukemia inhibitory factor / small-subunit processome / protein kinase C binding / positive regulation of apoptotic signaling pathway / positive regulation of protein-containing complex assembly / placenta development / spindle / Regulation of expression of SLITs and ROBOs / cytoplasmic ribonucleoprotein granule / modification-dependent protein catabolic process / G1/S transition of mitotic cell cycle / rRNA processing / protein tag activity / ribosomal small subunit biogenesis / rhythmic process / positive regulation of canonical Wnt signaling pathway / small ribosomal subunit rRNA binding / 加水分解酵素; 酸無水物に作用; GTPに作用・細胞または細胞小器官の運動に関与 / ribosome binding / glucose homeostasis / regulation of translation / ribosomal small subunit assembly / virus receptor activity / small ribosomal subunit / T cell differentiation in thymus / cell body / cytosolic small ribosomal subunit / perikaryon / cytoplasmic translation / mitochondrial inner membrane / cell differentiation / postsynaptic density / rRNA binding / ribosome / protein ubiquitination / structural constituent of ribosome / positive regulation of apoptotic process / ribonucleoprotein complex / positive regulation of protein phosphorylation / translation / cell division / DNA repair / GTPase activity / mRNA binding / centrosome / ubiquitin protein ligase binding / dendrite / positive regulation of cell population proliferation / synapse / GTP binding / negative regulation of apoptotic process / nucleolus / apoptotic process / protein kinase binding / perinuclear region of cytoplasm / Golgi apparatus / endoplasmic reticulum / DNA binding / RNA binding / zinc ion binding 類似検索 - 分子機能 | |||||||||
生物種 | HOMO SAPIENS (ヒト) ORYCTOLAGUS CUNICULUS (ウサギ) CRICKET PARALYSIS VIRUS (ウイルス) | |||||||||
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: 9 Å | |||||||||
データ登録者 | Muhs, M. / Hilal, T. / Mielke, T. / Skabkin, M.A. / Sanbonmatsu, K.Y. / Pestova, T.V. / Spahn, C.M.T. | |||||||||
引用 | ジャーナル: Mol Cell / 年: 2015 タイトル: Cryo-EM of ribosomal 80S complexes with termination factors reveals the translocated cricket paralysis virus IRES. 著者: Margarita Muhs / Tarek Hilal / Thorsten Mielke / Maxim A Skabkin / Karissa Y Sanbonmatsu / Tatyana V Pestova / Christian M T Spahn / 要旨: The cricket paralysis virus (CrPV) uses an internal ribosomal entry site (IRES) to hijack the ribosome. In a remarkable RNA-based mechanism involving neither initiation factor nor initiator tRNA, the ...The cricket paralysis virus (CrPV) uses an internal ribosomal entry site (IRES) to hijack the ribosome. In a remarkable RNA-based mechanism involving neither initiation factor nor initiator tRNA, the CrPV IRES jumpstarts translation in the elongation phase from the ribosomal A site. Here, we present cryoelectron microscopy (cryo-EM) maps of 80S⋅CrPV-STOP ⋅ eRF1 ⋅ eRF3 ⋅ GMPPNP and 80S⋅CrPV-STOP ⋅ eRF1 complexes, revealing a previously unseen binding state of the IRES and directly rationalizing that an eEF2-dependent translocation of the IRES is required to allow the first A-site occupation. During this unusual translocation event, the IRES undergoes a pronounced conformational change to a more stretched conformation. At the same time, our structural analysis provides information about the binding modes of eRF1 ⋅ eRF3 ⋅ GMPPNP and eRF1 in a minimal system. It shows that neither eRF3 nor ABCE1 are required for the active conformation of eRF1 at the intersection between eukaryotic termination and recycling. | |||||||||
履歴 |
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Remark 700 | SHEET DETERMINATION METHOD: DSSP THE SHEETS PRESENTED AS "BB" IN EACH CHAIN ON SHEET RECORDS BELOW ... SHEET DETERMINATION METHOD: DSSP THE SHEETS PRESENTED AS "BB" IN EACH CHAIN ON SHEET RECORDS BELOW IS ACTUALLY AN -7-STRANDED BARREL THIS IS REPRESENTED BY A -6-STRANDED SHEET IN WHICH THE FIRST AND LAST STRANDS ARE IDENTICAL. THE SHEETS PRESENTED AS "BD" IN EACH CHAIN ON SHEET RECORDS BELOW IS ACTUALLY AN -2-STRANDED BARREL THIS IS REPRESENTED BY A -1-STRANDED SHEET IN WHICH THE FIRST AND LAST STRANDS ARE IDENTICAL. |
-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 | 4d61.cif.gz | 1.9 MB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb4d61.ent.gz | 1.5 MB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 4d61.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 | 4d61_validation.pdf.gz | 1.1 MB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 4d61_full_validation.pdf.gz | 1.2 MB | 表示 | |
XML形式データ | 4d61_validation.xml.gz | 153.1 KB | 表示 | |
CIF形式データ | 4d61_validation.cif.gz | 264.5 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/d6/4d61 ftp://data.pdbj.org/pub/pdb/validation_reports/d6/4d61 | 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 , 2種, 2分子 1j
#1: RNA鎖 | 分子量: 602776.875 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ORYCTOLAGUS CUNICULUS (ウサギ) |
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#37: RNA鎖 | 分子量: 64363.910 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) CRICKET PARALYSIS VIRUS (ウイルス) / 参照: GenBank: AF218039 |
+40S RIBOSOMAL PROTEIN ... , 31種, 31分子 ABCDEFGHIJKLMNOPQRSTUVWXYZabcde
-タンパク質 , 2種, 2分子 fg
#33: タンパク質 | 分子量: 18004.041 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ORYCTOLAGUS CUNICULUS (ウサギ) / 参照: UniProt: G1SK22*PLUS |
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#34: タンパク質 | 分子量: 35115.652 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ORYCTOLAGUS CUNICULUS (ウサギ) / 参照: UniProt: G1SJB4*PLUS |
-EUKARYOTIC PEPTIDE CHAIN RELEASE FACTOR ... , 2種, 2分子 hi
#35: タンパク質 | 分子量: 49040.711 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) HOMO SAPIENS (ヒト) / 発現宿主: ESCHERICHIA COLI (大腸菌) / 株 (発現宿主): BL21(DE3) / 参照: UniProt: P62495 |
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#36: タンパク質 | 分子量: 47888.246 Da / 分子数: 1 / Fragment: G-DOMAIN, UNP RESIDUES 210-635 / 由来タイプ: 組換発現 / 由来: (組換発現) HOMO SAPIENS (ヒト) / 発現宿主: ESCHERICHIA COLI (大腸菌) / 株 (発現宿主): BL21(DE3) / 参照: UniProt: P15170 |
-詳細
配列の詳細 | AUTHORS HAVE USED HUMAN SEQUENCE FOR MODEL BUILDING, WITH FOLLOWING PDB-GENBANK OR UNIPROT RESIDUE ...AUTHORS HAVE USED HUMAN SEQUENCE FOR MODEL BUILDING, WITH FOLLOWING PDB-GENBANK OR UNIPROT RESIDUE MAPPING: CHAIN: 1 1-1742 GB X03205.1 1 1742 CHAIN: A 1-295 UNP P08865 1 295 CHAIN: B 1- 264 UNP P61247 1 264 CHAIN: C 1-293 UNP P15880 1 293 CHAIN: D 1-243 UNP P23396 1 243 CHAIN: E 1-263 UNP P22090 1 263 CHAIN: F 1-204 UNP P46782 1 204 CHAIN: G 1-249 UNP P62753 1 249 CHAIN: H 1-194 UNP P62081 1 194 CHAIN: I 1-208 UNP P62241 1 208 CHAIN: J 1-194 UNP P46781 1 194 CHAIN: K 1-165 UNP P46783 1 165 CHAIN: L 1-158 UNP P62280 1 158 CHAIN: M 1-132 UNP P25398 1 132 CHAIN: N 1-151 UNP P62277 1 151 CHAIN: O 1-151 UNP P62263 1 151 CHAIN: P 1-145 UNP P62841 1 145 CHAIN: Q 1-146 UNP P62249 1 146 CHAIN: R 1-135 UNP P08708 1 135 CHAIN: S 1-152 UNP P62269 1 152 CHAIN: T 1-145 UNP P39019 1 145 CHAIN: U 1-119 UNP P60866 1 119 CHAIN: V 1- 83 UNP P63220 1 83 CHAIN: W 1-130 UNP P62244 1 130 CHAIN: X 1-143 UNP P62266 1 143 CHAIN: Y 1-133 UNP P62847 1 133 CHAIN: Z 1-125 UNP P62851 1 125 CHAIN: a 1-115 UNP P62854 1 115 CHAIN: b 1-84 UNP P42677 1 84 CHAIN: c 1-69 UNP P62857 1 69 CHAIN: d 1-56 UNP P62273 1 56 CHAIN: e 1-59 UNP P62861 1 59 CHAIN: f 1-156 UNP P62979 1 156 CHAIN: g 1-317 UNP P63244 1 317 |
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-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: CRICKET PARALYSIS VIRUS IRES RNA BOUND TO MAMMALIAN 80S RIBOSOME, ERF1 AND ERF3 タイプ: RIBOSOME / 詳細: MICROGRAPHS WERE SELECTED FOR DRIFT AND ASTIGMATISM |
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緩衝液 | 名称: 20 MM TRIS PH 7.5, 100 MM KCL, 1 MM DTT, 8.5 MM MGCL2, 0.133 MM GTP, 2.33 MM GMPPNP pH: 7.5 詳細: 20 MM TRIS PH 7.5, 100 MM KCL, 1 MM DTT, 8.5 MM MGCL2, 0.133 MM GTP, 2.33 MM GMPPNP |
試料 | 濃度: 1.38 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: FEI VITROBOT MARK II / 凍結剤: ETHANE / 詳細: LIQUID ETHANE |
-電子顕微鏡撮影
実験機器 | モデル: Tecnai F20 / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TECNAI F20 / 日付: 2012年11月5日 / 詳細: MICROGRAPHS WERE SELECTED FOR DRIFT AND ASTIGMATISM |
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): 115000 X / 倍率(補正後): 194805 X / 最大 デフォーカス(公称値): 5000 nm / 最小 デフォーカス(公称値): 2000 nm / Cs: 2 mm |
試料ホルダ | 温度: 77 K |
撮影 | 電子線照射量: 20 e/Å2 フィルム・検出器のモデル: TVIPS TEMCAM-F416 (4k x 4k) |
画像スキャン | デジタル画像の数: 1 |
放射波長 | 相対比: 1 |
-解析
EMソフトウェア |
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CTF補正 | 詳細: DEFOCUS GROUPS | ||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||
3次元再構成 | 手法: MULTI-REFERENCE TEMPLATE MATCHING / 解像度: 9 Å / 粒子像の数: 64902 / ピクセルサイズ(公称値): 1.56 Å / ピクセルサイズ(実測値): 1.56 Å 倍率補正: CROSS- -CORRELATION DENSITIES WITH REFERENCE STRUCTURE 対称性のタイプ: POINT | ||||||||||||
原子モデル構築 | プロトコル: FLEXIBLE FIT / 空間: REAL / 詳細: METHOD--RIGID BODY, FLEXIBLE FIT | ||||||||||||
精密化 | 最高解像度: 9 Å | ||||||||||||
精密化ステップ | サイクル: LAST / 最高解像度: 9 Å
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