+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 6bu8 | |||||||||
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タイトル | 70S ribosome with S1 domains 1 and 2 (Class 1) | |||||||||
要素 |
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キーワード | RIBOSOME / tRNA / S1 / ribosomal protein S1 | |||||||||
機能・相同性 | 機能・相同性情報 RNA secondary structure unwinding / positive regulation of cytoplasmic translation / negative regulation of cytoplasmic translational initiation / stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation ...RNA secondary structure unwinding / positive regulation of cytoplasmic translation / negative regulation of cytoplasmic translational initiation / stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / positive regulation of ribosome biogenesis / negative regulation of cytoplasmic translation / four-way junction DNA binding / translational termination / DnaA-L2 complex / translation repressor activity / negative regulation of DNA-templated DNA replication initiation / negative regulation of translational initiation / regulation of mRNA stability / mRNA regulatory element binding translation repressor activity / ribosome assembly / assembly of large subunit precursor of preribosome / positive regulation of RNA splicing / transcription elongation factor complex / cytosolic ribosome assembly / regulation of DNA-templated transcription elongation / DNA endonuclease activity / response to reactive oxygen species / transcription antitermination / regulation of cell growth / translational initiation / DNA-templated transcription termination / maintenance of translational fidelity / response to radiation / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / ribosome biogenesis / ribosome binding / regulation of translation / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / transferase activity / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / tRNA binding / molecular adaptor activity / negative regulation of translation / single-stranded RNA binding / rRNA binding / ribosome / structural constituent of ribosome / translation / response to antibiotic / negative regulation of DNA-templated transcription / mRNA binding / DNA binding / RNA binding / zinc ion binding / membrane / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | Escherichia coli (大腸菌) | |||||||||
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: 3.5 Å | |||||||||
データ登録者 | Loveland, A.B. / Korostelev, A.A. | |||||||||
資金援助 | 米国, 2件
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引用 | ジャーナル: Methods / 年: 2018 タイトル: Structural dynamics of protein S1 on the 70S ribosome visualized by ensemble cryo-EM. 著者: Anna B Loveland / Andrei A Korostelev / 要旨: Bacterial ribosomal protein S1 is the largest and highly flexible protein of the 30S subunit, and one of a few core ribosomal proteins for which a complete structure is lacking. S1 is thought to ...Bacterial ribosomal protein S1 is the largest and highly flexible protein of the 30S subunit, and one of a few core ribosomal proteins for which a complete structure is lacking. S1 is thought to participate in transcription and translation. Best understood is the role of S1 in facilitating translation of mRNAs with structured 5' UTRs. Here, we present cryo-EM analyses of the 70S ribosome that reveal multiple conformations of S1. Based on comparison of several 3D maximum likelihood classification approaches in Frealign, we propose a streamlined strategy for visualizing a highly dynamic component of a large macromolecular assembly that itself exhibits high compositional and conformational heterogeneity. The resulting maps show how S1 docks at the ribosomal protein S2 near the mRNA exit channel. The globular OB-fold domains sample a wide area around the mRNA exit channel and interact with mobile tails of proteins S6 and S18. S1 also interacts with the mRNA entrance channel, where an OB-fold domain can be localized near S3 and S5. Our analyses suggest that S1 cooperates with other ribosomal proteins to form a dynamic mesh near the mRNA exit and entrance channels to modulate the binding, folding and movement of mRNA. | |||||||||
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 | 6bu8.cif.gz | 3.8 MB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb6bu8.ent.gz | 表示 | PDB形式 | |
PDBx/mmJSON形式 | 6bu8.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 | 6bu8_validation.pdf.gz | 1.3 MB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 6bu8_full_validation.pdf.gz | 1.7 MB | 表示 | |
XML形式データ | 6bu8_validation.xml.gz | 264.2 KB | 表示 | |
CIF形式データ | 6bu8_validation.cif.gz | 432.8 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/bu/6bu8 ftp://data.pdbj.org/pub/pdb/validation_reports/bu/6bu8 | 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
+50S ribosomal protein ... , 32種, 32分子 0405060708091011121314151617181920212223242526272829303132333403
+30S ribosomal protein ... , 21種, 21分子 BCDEFGHIJKLMNOPQRSTUZ
-RNA鎖 , 6種, 7分子 A0102XWVY
#53: RNA鎖 | 分子量: 498725.406 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Escherichia coli (strain K12) (大腸菌) / 株: K12 / 参照: GenBank: 1126835768 | ||||
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#54: RNA鎖 | 分子量: 941305.250 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Escherichia coli (strain K12) (大腸菌) / 株: K12 / 参照: GenBank: 802133627 | ||||
#55: RNA鎖 | 分子量: 38813.133 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Escherichia coli (strain K12) (大腸菌) / 株: K12 / 参照: GenBank: 1266961702 | ||||
#56: RNA鎖 | 分子量: 24802.785 Da / 分子数: 2 / 由来タイプ: 天然 / 由来: (天然) Escherichia coli (strain K12) (大腸菌) / 株: K12 / 参照: GenBank: 1275521620 #57: RNA鎖 | | 分子量: 8815.392 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Escherichia coli (strain K12) (大腸菌) / 株: K12 #58: RNA鎖 | | 分子量: 24485.539 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Escherichia coli (strain K12) (大腸菌) / 株: K12 / 参照: GenBank: 1275521620 |
-非ポリマー , 1種, 1分子
#60: 化合物 | ChemComp-FME / |
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-詳細
配列の詳細 | THE RESIDUE IDENTITY IS BASED ON HIGHER RESOLUTION |
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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: 70S ribosome with S1 domains 1 and 2 (Class 1) / タイプ: RIBOSOME / Entity ID: #1-#59 / 由来: NATURAL |
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分子量 | 値: 2.5 MDa / 実験値: NO |
由来(天然) | 生物種: Escherichia coli (strain K12) (大腸菌) / 株: K12 |
緩衝液 | pH: 7.5 |
試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES 詳細: 250 nM 50S, 250 nM 30S, 1.25 micromolar mRNA, 500 nM fMet-tRNAfMet, 1 micromolar EF-T, 500 micromolar GDPCP, 1 micromolar Phe-tRNAPhe |
試料支持 | 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: C-flat-1.2/1.3 4C |
急速凍結 | 装置: GATAN CRYOPLUNGE 3 / 凍結剤: ETHANE / 湿度: 90 % / 凍結前の試料温度: 275 K 詳細: 2 uL of complex was applied to each grid. After a 10-second incubation, the grids were blotted for 2 to 4 seconds. |
-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 / 倍率(公称値): 60976 X / 倍率(補正後): 60976 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): 500 nm / Cs: 2.7 mm / 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: 0.4 sec. / 電子線照射量: 1 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) 撮影したグリッド数: 2 / 実像数: 3928 |
画像スキャン | サンプリングサイズ: 5 µm / 横: 7676 / 縦: 7420 / 動画フレーム数/画像: 50 / 利用したフレーム数/画像: 1-50 |
-解析
EMソフトウェア |
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画像処理 | 詳細: Gain reference was applied, movies were aligned, and the summed imaged were corrected for magnification anisotropy. | ||||||||||||||||||||||||||||||||||||||||||||||||
CTF補正 | 詳細: CTFFIND3 was used to determine CTF values. FREALIGN applied CTF correction. タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||||||||||
粒子像の選択 | 選択した粒子像数: 800367 詳細: Particles were picked from micrographs using Signature reference-based particle picker. | ||||||||||||||||||||||||||||||||||||||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||||||||||||||||||||||||||||||||||||||
3次元再構成 | 解像度: 3.5 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 10289 / アルゴリズム: BACK PROJECTION / クラス平均像の数: 16 / 対称性のタイプ: POINT | ||||||||||||||||||||||||||||||||||||||||||||||||
原子モデル構築 | プロトコル: RIGID BODY FIT / 空間: REAL / Target criteria: correlation coefficient |