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データを開く
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基本情報
| 登録情報 | データベース: PDB / ID: 7r81 | ||||||||||||
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| タイトル | Structure of the translating Neurospora crassa ribosome arrested by cycloheximide | ||||||||||||
要素 |
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キーワード | TRANSLATION / Protein Synthesis Inhibitor / Cytosolic Ribosome / Polysomal Ribosome / Peptidyl Transferase Center | ||||||||||||
| 機能・相同性 | 機能・相同性情報negative regulation of cell integrity MAPK cascade / positive regulation of conjugation with cellular fusion / regulation of cytoplasmic translation / GCN2-mediated signaling / negative regulation of p38MAPK cascade / ribosome hibernation / preribosome / regulation of amino acid metabolic process / negative regulation of glucose mediated signaling pathway / positive regulation of translational fidelity ...negative regulation of cell integrity MAPK cascade / positive regulation of conjugation with cellular fusion / regulation of cytoplasmic translation / GCN2-mediated signaling / negative regulation of p38MAPK cascade / ribosome hibernation / preribosome / regulation of amino acid metabolic process / negative regulation of glucose mediated signaling pathway / positive regulation of translational fidelity / ribosome-associated ubiquitin-dependent protein catabolic process / pre-mRNA 5'-splice site binding / GDP-dissociation inhibitor activity / nonfunctional rRNA decay / preribosome, small subunit precursor / mRNA destabilization / negative regulation of mRNA splicing, via spliceosome / signaling receptor activator activity / negative regulation of translational frameshifting / translational elongation / G-protein alpha-subunit binding / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / ribosomal subunit export from nucleus / protein-RNA complex assembly / translation regulator activity / 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) / protein-membrane adaptor activity / ribosomal small subunit export from nucleus / DNA-(apurinic or apyrimidinic site) endonuclease activity / positive regulation of autophagy / rescue of stalled cytosolic ribosome / protein kinase C binding / ribosomal large subunit biogenesis / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / maintenance of translational fidelity / modification-dependent protein catabolic process / protein tag activity / rRNA processing / large ribosomal subunit / ribosomal small subunit assembly / ribosome biogenesis / ribosome binding / ribosomal small subunit biogenesis / 5S rRNA binding / small ribosomal subunit / ribosomal large subunit assembly / small ribosomal subunit rRNA binding / cytosolic small ribosomal subunit / large ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / cytoplasmic translation / rRNA binding / structural constituent of ribosome / protein ubiquitination / ribosome / translation / G protein-coupled receptor signaling pathway / ribonucleoprotein complex / mRNA binding / ubiquitin protein ligase binding / nucleolus / RNA binding / zinc ion binding / nucleus / cytoplasm / cytosol 類似検索 - 分子機能 | ||||||||||||
| 生物種 | Neurospora crassa (菌類) | ||||||||||||
| 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: 2.7 Å | ||||||||||||
データ登録者 | Shen, L. / Su, Z. / Yang, K. / Wu, C. / Becker, T. / Bell-Pedersen, D. / Zhang, J. / Sachs, M.S. | ||||||||||||
| 資金援助 | 米国, 3件
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引用 | ジャーナル: Proc Natl Acad Sci U S A / 年: 2021タイトル: Structure of the translating ribosome arrested by cycloheximide. 著者: Lunda Shen / Zhaoming Su / Kailu Yang / Cheng Wu / Thomas Becker / Deborah Bell-Pedersen / Junjie Zhang / Matthew S Sachs / ![]() 要旨: Ribosomes translate RNA into proteins. The protein synthesis inhibitor cycloheximide (CHX) is widely used to inhibit eukaryotic ribosomes engaged in translation elongation. However, the lack of ...Ribosomes translate RNA into proteins. The protein synthesis inhibitor cycloheximide (CHX) is widely used to inhibit eukaryotic ribosomes engaged in translation elongation. However, the lack of structural data for actively translating polyribosomes stalled by CHX leaves unanswered the question of which elongation step is inhibited. We elucidated CHX's mechanism of action based on the cryo-electron microscopy structure of actively translating ribosomes bound with CHX at 2.7-Å resolution. The ribosome structure from this filamentous fungus contains clearly resolved ribosomal protein eL28, like higher eukaryotes but unlike budding yeast, which lacks eL28. Despite some differences in overall structures, the ribosomes from , yeast, and humans all contain a highly conserved CHX binding site. We also sequenced classic CHX-resistant alleles. These mutations, including one at a residue not previously observed to affect CHX resistance in eukaryotes, were in the large subunit proteins uL15 and eL42 that are part of the CHX-binding pocket. In addition to A-site transfer RNA (tRNA), P-site tRNA, messenger RNA, and CHX that are associated with the translating ribosome, spermidine is present near the CHX binding site close to the E site on the large subunit. The tRNAs in the peptidyl transferase center are in the A/A site and the P/P site. The nascent peptide is attached to the A-site tRNA and not to the P-site tRNA. The structural and functional data obtained show that CHX arrests the ribosome in the classical PRE translocation state and does not interfere with A-site reactivity. | ||||||||||||
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構造の表示
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ムービービューア |
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| 構造ビューア | 分子: Molmil Jmol/JSmol |
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ダウンロードとリンク
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ダウンロード
| PDBx/mmCIF形式 | 7r81.cif.gz | 4.4 MB | 表示 | PDBx/mmCIF形式 |
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| PDB形式 | pdb7r81.ent.gz | 表示 | PDB形式 | |
| PDBx/mmJSON形式 | 7r81.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
| アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/r8/7r81 ftp://data.pdbj.org/pub/pdb/validation_reports/r8/7r81 | HTTPS FTP |
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-関連構造データ
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リンク
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集合体
| 登録構造単位 | ![]()
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要素
-RNA鎖 , 6種, 7分子 A1A2B1C1t1u1w1
| #1: RNA鎖 | 分子量: 1078431.750 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: GenBank: 209980056 | ||
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| #2: RNA鎖 | 分子量: 578821.625 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: GenBank: 209980056 | ||
| #3: RNA鎖 | 分子量: 38579.906 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: GenBank: 168880 | ||
| #5: RNA鎖 | 分子量: 50770.008 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: GenBank: 168899 | ||
| #80: RNA鎖 | 分子量: 24468.551 Da / 分子数: 2 / 由来タイプ: 天然 詳細: One molecule is found in the A-site, and one molecule is found in the P-site. No specific tRNAs are represented in the peptidyltransferase centers of polysomal ribosomes. Therefore, we ...詳細: One molecule is found in the A-site, and one molecule is found in the P-site. No specific tRNAs are represented in the peptidyltransferase centers of polysomal ribosomes. Therefore, we adapted the tRNA model for a eukaryotic ICG-anticodon tRNA that decodes Arg codons (PDB ID: 6T4Q) to model the tRNA densities of both A- and P-site tRNAs. 由来: (天然) Neurospora crassa (菌類)#82: RNA鎖 | | 分子量: 5159.861 Da / 分子数: 1 / 由来タイプ: 天然 詳細: No specific mRNA is represented in the polysomal ribosomes. Therefore, we used Uracil to model the mRNA. 由来: (天然) Neurospora crassa (菌類) |
+40S ribosomal protein ... , 28種, 28分子 B2C2D2F2G2H2I2J2K2M2N2O2P2R2S2T2U2V2W2X2Y2Z2a2b2c2d2e2f2
+60S ribosomal protein ... , 34種, 34分子 D1E1F1G1H1I1J1K1L1M1N1P1Q1R1S1U1W1X1Y1Z1b1c1d1e1g1h1i1j1k1m1...
-タンパク質 , 6種, 6分子 E2L2f1g2h2s1
| #10: タンパク質 | 分子量: 28706.771 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: UniProt: Q7RV52 |
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| #24: タンパク質 | 分子量: 18463.754 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: UniProt: Q873A6 |
| #63: タンパク質 | 分子量: 14097.444 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: UniProt: Q7RW15 |
| #66: タンパク質 | 分子量: 17775.771 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: UniProt: P14799 |
| #68: タンパク質 | 分子量: 35173.578 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: UniProt: Q01369 |
| #79: タンパク質 | 分子量: 16011.481 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: UniProt: Q7SBB3 |
-Ribosomal protein ... , 6種, 6分子 O1Q2T1V1a1l1
| #29: タンパク質 | 分子量: 15885.671 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: UniProt: Q7SI18 |
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| #34: タンパク質 | 分子量: 17419.408 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: UniProt: Q7SGF1 |
| #39: タンパク質 | 分子量: 22407.271 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: UniProt: Q7S5N9 |
| #43: タンパク質 | 分子量: 18253.395 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: UniProt: Q7SDY8 |
| #53: タンパク質 | 分子量: 15366.948 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: UniProt: Q1K4V7 |
| #72: タンパク質 | 分子量: 10261.002 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: UniProt: V5IQ48 |
-タンパク質・ペプチド , 2種, 2分子 p1v1
| #76: タンパク質・ペプチド | 分子量: 3384.335 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Neurospora crassa (菌類) / 参照: UniProt: U9WGG3 |
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| #81: タンパク質・ペプチド | 分子量: 358.434 Da / 分子数: 1 / 由来タイプ: 天然 詳細: No specific nascent peptide is represented in the polysomal ribosomes. Therefore, we used UNK (unknown residue) to represent each residue of the nascent peptide. 由来: (天然) Neurospora crassa (菌類) |
-非ポリマー , 3種, 262分子 




| #83: 化合物 | ChemComp-3HE / |
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| #84: 化合物 | ChemComp-SPD / |
| #85: 化合物 | ChemComp-MG / |
-詳細
| 研究の焦点であるリガンドがあるか | Y |
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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 |
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試料調製
| 構成要素 |
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| 由来(天然) |
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| 緩衝液 | pH: 7.6 | ||||||||||||||||||||||||||||||||||||||||||
| 試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | ||||||||||||||||||||||||||||||||||||||||||
| 急速凍結 | 凍結剤: ETHANE |
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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 |
| 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 |
| 撮影 | 電子線照射量: 32 e/Å2 / 検出モード: SUPER-RESOLUTION フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) |
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解析
| ソフトウェア | 名称: UCSF ChimeraX / バージョン: 1.1/v9 / 分類: モデル構築 / URL: https://www.rbvi.ucsf.edu/chimerax/ / Os: macOS / タイプ: package |
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| CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION |
| 3次元再構成 | 解像度: 2.7 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 196154 / 対称性のタイプ: POINT |
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コントローラー
万見について




Neurospora crassa (菌類)
米国, 3件
引用
UCSF Chimera










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