+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 6h4n | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
タイトル | Structure of a hibernating 100S ribosome reveals an inactive conformation of the ribosomal protein S1 - 70S Hibernating E. coli Ribosome | |||||||||
要素 |
| |||||||||
キーワード | RIBOSOME / 100S / cryo-EM / E-site tRNA / hibernation / HPF / RMF / S1 | |||||||||
機能・相同性 | 機能・相同性情報 negative regulation of translation in response to stress / dormancy process / negative regulation of translational elongation / RNA secondary structure unwinding / positive regulation of cytoplasmic translation / negative regulation of cytoplasmic translational initiation / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing ...negative regulation of translation in response to stress / dormancy process / negative regulation of translational elongation / RNA secondary structure unwinding / positive regulation of cytoplasmic translation / negative regulation of cytoplasmic translational initiation / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / ribosomal small subunit binding / 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 / DnaA-L2 complex / translation repressor activity / negative regulation of translational initiation / negative regulation of DNA-templated DNA replication initiation / regulation of mRNA stability / mRNA regulatory element binding translation repressor activity / ribosome assembly / positive regulation of RNA splicing / assembly of large subunit precursor of preribosome / transcription elongation factor complex / cytosolic ribosome assembly / regulation of DNA-templated transcription elongation / DNA endonuclease activity / ribosomal large subunit assembly / transcription antitermination / response to reactive oxygen species / translational initiation / regulation of cell growth / DNA-templated transcription termination / maintenance of translational fidelity / response to radiation / mRNA 5'-UTR binding / large ribosomal subunit / ribosome biogenesis / ribosome binding / regulation of translation / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / small ribosomal subunit rRNA binding / transferase activity / 5S rRNA binding / large ribosomal subunit rRNA binding / cytosolic small ribosomal subunit / cytosolic large ribosomal subunit / cytoplasmic translation / tRNA binding / molecular adaptor activity / single-stranded RNA binding / rRNA binding / negative regulation of translation / 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 / cytoplasm / cytosol 類似検索 - 分子機能 | |||||||||
生物種 | Escherichia coli BW25113 (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 Å | |||||||||
データ登録者 | Beckert, B. / Turk, M. / Czech, A. / Berninghausen, O. / Beckmann, R. / Ignatova, Z. / Plitzko, J. / Wilson, N.D. | |||||||||
資金援助 | ドイツ, 2件
| |||||||||
引用 | ジャーナル: Nat Microbiol / 年: 2018 タイトル: Structure of a hibernating 100S ribosome reveals an inactive conformation of the ribosomal protein S1. 著者: Bertrand Beckert / Martin Turk / Andreas Czech / Otto Berninghausen / Roland Beckmann / Zoya Ignatova / Jürgen M Plitzko / Daniel N Wilson / 要旨: To survive under conditions of stress, such as nutrient deprivation, bacterial 70S ribosomes dimerize to form hibernating 100S particles. In γ-proteobacteria, such as Escherichia coli, 100S ...To survive under conditions of stress, such as nutrient deprivation, bacterial 70S ribosomes dimerize to form hibernating 100S particles. In γ-proteobacteria, such as Escherichia coli, 100S formation requires the ribosome modulation factor (RMF) and the hibernation promoting factor (HPF). Here we present single-particle cryo-electron microscopy structures of hibernating 70S and 100S particles isolated from stationary-phase E. coli cells at 3.0 Å and 7.9 Å resolution, respectively. The structures reveal the binding sites for HPF and RMF as well as the unexpected presence of deacylated E-site transfer RNA and ribosomal protein bS1. HPF interacts with the anticodon-stem-loop of the E-tRNA and occludes the binding site for the messenger RNA as well as A- and P-site tRNAs. RMF facilitates stabilization of a compact conformation of bS1, which together sequester the anti-Shine-Dalgarno sequence of the 16S ribosomal RNA (rRNA), thereby inhibiting translation initiation. At the dimerization interface, the C-terminus of uS2 probes the mRNA entrance channel of the symmetry-related particle, thus suggesting that dimerization inactivates ribosomes by blocking the binding of mRNA within the channel. The back-to-back E. coli 100S arrangement is distinct from 100S particles observed previously in Gram-positive bacteria, and reveals a unique role for bS1 in translation regulation. | |||||||||
履歴 |
|
-構造の表示
ムービー |
ムービービューア |
---|---|
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 | 6h4n.cif.gz | 3.2 MB | 表示 | PDBx/mmCIF形式 |
---|---|---|---|---|
PDB形式 | pdb6h4n.ent.gz | 表示 | PDB形式 | |
PDBx/mmJSON形式 | 6h4n.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 | 6h4n_validation.pdf.gz | 1.2 MB | 表示 | wwPDB検証レポート |
---|---|---|---|---|
文書・詳細版 | 6h4n_full_validation.pdf.gz | 1.3 MB | 表示 | |
XML形式データ | 6h4n_validation.xml.gz | 213.8 KB | 表示 | |
CIF形式データ | 6h4n_validation.cif.gz | 377 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/h4/6h4n ftp://data.pdbj.org/pub/pdb/validation_reports/h4/6h4n | HTTPS FTP |
-関連構造データ
-リンク
-集合体
登録構造単位 |
|
---|---|
1 |
|
-要素
-RNA鎖 , 5種, 5分子 aABwI
#1: RNA鎖 | 分子量: 497197.531 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Escherichia coli BW25113 (大腸菌) / 参照: GenBank: 1359176725 |
---|---|
#22: 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 BW25113 (大腸菌) / 参照: GenBank: 1036415628 |
#23: 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 BW25113 (大腸菌) / 参照: GenBank: 1393501787 |
#53: RNA鎖 | 分子量: 24771.770 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Escherichia coli BW25113 (大腸菌) / 参照: GenBank: 1063812051 |
#57: RNA鎖 | 分子量: 1482.920 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Escherichia coli BW25113 (大腸菌) |
+30S ribosomal protein ... , 21種, 21分子 bcdefghijklmnopqrstuy
+50S ribosomal protein ... , 29種, 29分子 CDEFGHJKLMNOPQRSTUVWXYZ012346
-タンパク質 , 2種, 2分子 vx
#54: タンパク質 | 分子量: 6520.523 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Escherichia coli BW25113 (大腸菌) / 参照: UniProt: P0AFW2 |
---|---|
#56: タンパク質 | 分子量: 10768.230 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Escherichia coli BW25113 (大腸菌) / 参照: UniProt: P0AFX0 |
-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
---|---|
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: Structure of a hibernating 100S ribosome reveals an inactive conformation of the ribosomal protein S1 タイプ: RIBOSOME 詳細: Here we present single particle cryo-electron microscopy structures of hibernating 70S and 100S particles isolated from stationary phase E. coli cells at 3.0 and 7.9 resolution, respectively. ...詳細: Here we present single particle cryo-electron microscopy structures of hibernating 70S and 100S particles isolated from stationary phase E. coli cells at 3.0 and 7.9 resolution, respectively. The structures reveal the binding sites for HPF and RMF as well as the unexpected presence of deacylated E-site tRNA and ribosomal protein bS1. HPF interacts with the anticodon-stem-loop of the E-tRNA and occludes the binding site for the mRNA as well as A- and P-site tRNAs. RMF facilitates stabilization of a compact conformation of bS1, which together sequester the anti-Shine-Dalgarno sequence of the 16S ribosomal RNA (rRNA), thereby inhibiting translation initiation. At the dimerization interface, the C-terminus of uS2 probes the mRNA entrance channel of the symmetry related particle, thus suggesting that dimerization inactivates ribosomes by blocking the binding of mRNA within the channel. Entity ID: all / 由来: NATURAL |
---|---|
分子量 | 単位: MEGADALTONS / 実験値: NO |
由来(天然) | 生物種: Escherichia coli BW25113 (大腸菌) |
緩衝液 | pH: 7.5 詳細: B100S buffer (25 mM Hepes pH 7.5, 100 mM KOAc, 15 mM Mg(OAc)2, 1 mM dithiothreitol) |
試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
試料支持 | グリッドの材料: GOLD / グリッドのタイプ: Quantifoil, UltrAuFoil, R1.2/1.3 |
急速凍結 | 装置: FEI VITROBOT MARK III / 凍結剤: ETHANE-PROPANE |
-電子顕微鏡撮影
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
---|---|
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: SPOT SCAN |
電子レンズ | モード: BRIGHT FIELD |
撮影 | 電子線照射量: 2.5 e/Å2 フィルム・検出器のモデル: FEI FALCON II (4k x 4k) |
-解析
EMソフトウェア |
| ||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||||||||||||||
3次元再構成 | 解像度: 3 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 188304 / 対称性のタイプ: POINT | ||||||||||||||||||||||||
原子モデル構築 | プロトコル: RIGID BODY FIT |