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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: 7qep | |||||||||||||||||||||
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タイトル | Cryo-EM structure of the ribosome from Encephalitozoon cuniculi | |||||||||||||||||||||
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![]() | RIBOSOME / genome decay / microbial parasite / genome reduction / Encephalitozoon cuniculi / lose-to-gain | |||||||||||||||||||||
機能・相同性 | ![]() rRNA processing / large ribosomal subunit / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / cytosolic small ribosomal subunit / cytosolic large ribosomal subunit / rRNA binding / ribosome / structural constituent of ribosome ...rRNA processing / large ribosomal subunit / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / cytosolic small ribosomal subunit / cytosolic large ribosomal subunit / rRNA binding / ribosome / structural constituent of ribosome / translation / nucleolus / RNA binding / nucleus / metal ion binding / cytoplasm 類似検索 - 分子機能 | |||||||||||||||||||||
生物種 | ![]() | |||||||||||||||||||||
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 | |||||||||||||||||||||
![]() | Nicholson, D. / Ranson, N.A. / Melnikov, S.V. | |||||||||||||||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Adaptation to genome decay in the structure of the smallest eukaryotic ribosome. 著者: David Nicholson / Marco Salamina / Johan Panek / Karla Helena-Bueno / Charlotte R Brown / Robert P Hirt / Neil A Ranson / Sergey V Melnikov / ![]() 要旨: The evolution of microbial parasites involves the counterplay between natural selection forcing parasites to improve and genetic drifts forcing parasites to lose genes and accumulate deleterious ...The evolution of microbial parasites involves the counterplay between natural selection forcing parasites to improve and genetic drifts forcing parasites to lose genes and accumulate deleterious mutations. Here, to understand how this counterplay occurs at the scale of individual macromolecules, we describe cryo-EM structure of ribosomes from Encephalitozoon cuniculi, a eukaryote with one of the smallest genomes in nature. The extreme rRNA reduction in E. cuniculi ribosomes is accompanied with unparalleled structural changes, such as the evolution of previously unknown molten rRNA linkers and bulgeless rRNA. Furthermore, E. cuniculi ribosomes withstand the loss of rRNA and protein segments by evolving an ability to use small molecules as structural mimics of degenerated rRNA and protein segments. Overall, we show that the molecular structures long viewed as reduced, degenerated, and suffering from debilitating mutations possess an array of compensatory mechanisms that allow them to remain active despite the extreme molecular reduction. | |||||||||||||||||||||
履歴 |
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構造の表示
ムービー |
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構造ビューア | 分子: ![]() ![]() |
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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 | ![]() | 3.6 MB | 表示 | ![]() |
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PDB形式 | ![]() | 表示 | ![]() | |
PDBx/mmJSON形式 | ![]() | ツリー表示 | ![]() | |
その他 | ![]() |
-検証レポート
文書・要旨 | ![]() | 1.5 MB | 表示 | ![]() |
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文書・詳細版 | ![]() | 1.8 MB | 表示 | |
XML形式データ | ![]() | 312.2 KB | 表示 | |
CIF形式データ | ![]() | 489.1 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-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
-タンパク質 , 8種, 8分子 RAC5D1M4M5MDMSP0
#1: タンパク質 | 分子量: 36828.836 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) ![]() 株: GB-M1 / 参照: UniProt: M1K775 |
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#10: タンパク質 | 分子量: 16383.311 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) ![]() 株: GB-M1 / 参照: UniProt: Q8SRE7 |
#16: タンパク質 | 分子量: 7661.769 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) ![]() 株: GB-M1 / 参照: UniProt: I7L8M6 |
#38: タンパク質 | 分子量: 12275.196 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) ![]() 株: GB-M1 / 参照: UniProt: I7L8J2 |
#39: タンパク質 | 分子量: 23823.924 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) ![]() 株: GB-M1 / 参照: UniProt: Q8SQU2 |
#44: タンパク質 | 分子量: 19453.312 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) ![]() 株: GB-M1 / 参照: UniProt: Q8SWQ4 |
#45: タンパク質 | 分子量: 9019.900 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) ![]() 株: GB-M1 / 参照: UniProt: I7IV41 |
#65: タンパク質 | 分子量: 14312.952 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) ![]() 株: GB-M1 / 参照: UniProt: Q8SRH8 |
-RNA鎖 , 3種, 3分子 123
#2: RNA鎖 | 分子量: 807085.000 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) ![]() 株: GB-M1 / 参照: GenBank: 13560063 |
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#3: RNA鎖 | 分子量: 38478.938 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) ![]() 株: GB-M1 / 参照: GenBank: 13560072 |
#4: RNA鎖 | 分子量: 422123.406 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) ![]() 株: GB-M1 / 参照: GenBank: 13560063 |
+40S RIBOSOMAL PROTEIN ... , 28種, 28分子 C0C1C2C3C4C6C7C8C9D0D2D3D4D5D6D7D8D9S0S1S2S3S4S5S6S7S8S9
-Similarity to ... , 2種, 2分子 E1N4
#25: タンパク質 | 分子量: 17141.877 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) ![]() 株: GB-M1 / 参照: UniProt: Q8SWB3 |
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#50: タンパク質 | 分子量: 11350.204 Da / 分子数: 1 / 由来タイプ: 天然 由来: (天然) ![]() 株: GB-M1 / 参照: UniProt: Q8SUZ9 |
+60S RIBOSOMAL PROTEIN ... , 36種, 36分子 L1L2L3L4L5L6L7L8L9M0M1M3M6M7M8M9N0N1N2N3N5N6N7N8N9O0O1O2O3O4...
-非ポリマー , 3種, 10分子 ![](data/chem/img/ZN.gif)
![](data/chem/img/AMP.gif)
![](data/chem/img/SPD.gif)
![](data/chem/img/AMP.gif)
![](data/chem/img/SPD.gif)
#78: 化合物 | ChemComp-ZN / #79: 化合物 | ChemComp-AMP / | #80: 化合物 | ChemComp-SPD / | |
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-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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分子量 |
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由来(天然) |
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緩衝液 | pH: 7.5 | ||||||||||||||||||||||||
緩衝液成分 |
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試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | ||||||||||||||||||||||||
試料支持 | 詳細: Quorum GloQube, 10 mA / グリッドの材料: COPPER / グリッドのサイズ: 400 divisions/in. / グリッドのタイプ: Quantifoil R1.2/1.3 | ||||||||||||||||||||||||
急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 277 K 詳細: Quantifoil grids (R1.2/1.3, 400 mesh, copper) were glow discharged (10 mA, 30s, Quorum GloQube), and 3 microlitres of the crude sample of E. cuniculi ribosomes (300 nM) was pipetted onto a ...詳細: Quantifoil grids (R1.2/1.3, 400 mesh, copper) were glow discharged (10 mA, 30s, Quorum GloQube), and 3 microlitres of the crude sample of E. cuniculi ribosomes (300 nM) was pipetted onto a grid. Excess sample was immediately blotted off and vitrification was performed by plunging the grid into liquid nitrogen-cooled liquid ethane at 100% humidity and 4 degrees celsius using an FEI Vitrobot Mark IV (Thermo Fisher) |
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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: ![]() |
電子レンズ | モード: BRIGHT FIELD / 倍率(公称値): 96000 X / 最大 デフォーカス(公称値): 2600 nm / 最小 デフォーカス(公称値): 800 nm / Cs: 2.7 mm / C2レンズ絞り径: 70 µm / アライメント法: COMA FREE |
試料ホルダ | 凍結剤: NITROGEN 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
撮影 | 平均露光時間: 1.35 sec. / 電子線照射量: 60 e/Å2 / 検出モード: INTEGRATING フィルム・検出器のモデル: FEI FALCON III (4k x 4k) 実像数: 2210 |
画像スキャン | 横: 4096 / 縦: 4096 |
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解析
ソフトウェア |
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EMソフトウェア |
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画像処理 | 詳細: Drift-corrected and dose-corrected averages of each movie were created using RELION 3.1s own implementation of motion correction and the contrast transfer functions estimated using CTFFIND-4.1 | ||||||||||||||||||||||||||||||||||||||||
CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||
粒子像の選択 | 選択した粒子像数: 260895 詳細: Particles were picked using Laplacian-of-Gaussian autopicking and reference-free 2D classification used to generate templates for further autopicking. The resulting particles were extracted ...詳細: Particles were picked using Laplacian-of-Gaussian autopicking and reference-free 2D classification used to generate templates for further autopicking. The resulting particles were extracted with binning-by-4, and 2D and 3D classification performed to remove junk images | ||||||||||||||||||||||||||||||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||||||||||||||||||||||||||||||
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: 108005 詳細: The remaining particles were re-extracted without binning and aligned and refined in 3D, again using a 60 Angstrom low-passed filtered ab initio starting model. Rounds of CTF refinement and ...詳細: The remaining particles were re-extracted without binning and aligned and refined in 3D, again using a 60 Angstrom low-passed filtered ab initio starting model. Rounds of CTF refinement and Bayesian polishing were performed until the map resolution stopped improving. 108,005 particles fed into the final 3D reconstruction of estimated resolution 2.7 Angstrom. クラス平均像の数: 1 / 対称性のタイプ: POINT | ||||||||||||||||||||||||||||||||||||||||
原子モデル構築 | プロトコル: OTHER / 空間: REAL / Target criteria: correlation coefficient 詳細: The model was built using fragments of S. cerevisiae (pdb id 4v88) and V. necatrix ribosomes (pdb id 6rm3) as starting models that were edited using Coot using genomic sequences of the E. ...詳細: The model was built using fragments of S. cerevisiae (pdb id 4v88) and V. necatrix ribosomes (pdb id 6rm3) as starting models that were edited using Coot using genomic sequences of the E. cuniculi strain GB-M1 to model rRNA and ribosomal proteins. For ribosomal proteins that are encoded by two alternative genes (with one gene coding for a zinc-coordinating protein and another gene coding for a zinc-free ribosomal protein), we used zinc-coordinating isoforms, because the cryo-EM map revealed the presence of these isoforms and not their zinc-free paralogs in the ribosome structure. The identity of protein msL2 in the ribosome structure was determined using the genomic sequence of the E. cuniculi strain GB-M1 and the cryo-EM map that revealed a unique combination of aromatic and bulky amino acids in its structure: the cryo-EM map showed that msL2 has a tyrosine residue at position 5, a tryptophan residue at position 9, and lysine or arginine residues at positions 10, 12 and 13. The only protein with this sequence was the hypothetical protein ECU06_1135, whose sequence and length were fully consistent with the cryo-EM map. The structure of E. cuniculi ribosomes was refined using Phenix real space refine and validated using MolProbity within Phenix and PDB OneDep. The parts of the model corresponding to the 60S, 40S body and 40S head were built and refined using the consensus map, 40S body multibody map and 40S head multibody map, respectively. | ||||||||||||||||||||||||||||||||||||||||
原子モデル構築 |
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精密化 | 立体化学のターゲット値: CDL v1.2 | ||||||||||||||||||||||||||||||||||||||||
拘束条件 |
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