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基本情報
登録情報 | データベース: EMDB / ID: EMD-8522 | |||||||||
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タイトル | 3.2 A cryo-EM ArfA-RF2 ribosome rescue complex (Structure I) | |||||||||
![]() | 3.2 A cryo-EM map of ArfA ribosome rescue complex (Structure I) | |||||||||
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![]() | ARFA RF2 ribosome rescue complex / RIBOSOME | |||||||||
機能・相同性 | ![]() translation release factor activity, codon specific / negative regulation of cytoplasmic translational initiation / stringent response / ribosomal large subunit binding / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation ...translation release factor activity, codon specific / negative regulation of cytoplasmic translational initiation / stringent response / ribosomal large subunit binding / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / positive regulation of ribosome biogenesis / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / translational termination / negative regulation of cytoplasmic translation / four-way junction DNA binding / DnaA-L2 complex / translation repressor activity / negative regulation of translational initiation / regulation of mRNA stability / negative regulation of DNA-templated DNA replication initiation / rescue of stalled ribosome / mRNA regulatory element binding translation repressor activity / assembly of large subunit precursor of preribosome / positive regulation of RNA splicing / ribosome assembly / transcription elongation factor complex / regulation of DNA-templated transcription elongation / cytosolic ribosome assembly / response to reactive oxygen species / DNA endonuclease activity / transcription antitermination / translational initiation / regulation of cell growth / DNA-templated transcription termination / response to radiation / maintenance of translational fidelity / mRNA 5'-UTR binding / regulation of translation / ribosome biogenesis / large ribosomal subunit / transferase activity / ribosome binding / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / ribosomal large subunit assembly / cytosolic small ribosomal subunit / large ribosomal subunit rRNA binding / small ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / cytoplasmic translation / tRNA binding / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / viral translational frameshifting / response to antibiotic / negative regulation of DNA-templated transcription / mRNA binding / DNA binding / RNA binding / zinc ion binding / metal ion binding / membrane / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | ![]() ![]() ![]() ![]() | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.2 Å | |||||||||
![]() | Demo G / Svidritskiy E | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Mechanism of ribosome rescue by ArfA and RF2. 著者: Gabriel Demo / Egor Svidritskiy / Rohini Madireddy / Ruben Diaz-Avalos / Timothy Grant / Nikolaus Grigorieff / Duncan Sousa / Andrei A Korostelev / ![]() 要旨: ArfA rescues ribosomes stalled on truncated mRNAs by recruiting release factor RF2, which normally binds stop codons to catalyze peptide release. We report two 3.2 Å resolution cryo-EM structures - ...ArfA rescues ribosomes stalled on truncated mRNAs by recruiting release factor RF2, which normally binds stop codons to catalyze peptide release. We report two 3.2 Å resolution cryo-EM structures - determined from a single sample - of the 70S ribosome with ArfA•RF2 in the A site. In both states, the ArfA C-terminus occupies the mRNA tunnel downstream of the A site. One state contains a compact inactive RF2 conformation. Ordering of the ArfA N-terminus in the second state rearranges RF2 into an extended conformation that docks the catalytic GGQ motif into the peptidyl-transferase center. Our work thus reveals the structural dynamics of ribosome rescue. The structures demonstrate how ArfA 'senses' the vacant mRNA tunnel and activates RF2 to mediate peptide release without a stop codon, allowing stalled ribosomes to be recycled. | |||||||||
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構造の表示
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構造ビューア | EMマップ: ![]() ![]() ![]() |
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画像 | ![]() | 87.3 KB | ||
Filedesc metadata | ![]() | 14.9 KB | ||
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EMDBのページ | ![]() ![]() |
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「今月の分子」の関連する項目 |
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注釈 | 3.2 A cryo-EM map of ArfA ribosome rescue complex (Structure I) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.215 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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試料の構成要素
+全体 : ArfA-RF2 ribosome rescue complex (Structure I)
+超分子 #1: ArfA-RF2 ribosome rescue complex (Structure I)
+分子 #1: 16S ribosomal RNA
+分子 #2: 23S ribosomal RNA
+分子 #3: 5S ribosomal RNA
+分子 #5: fMet-tRNA (P- and E-site)
+分子 #38: truncated mRNA
+分子 #4: Alternative ribosome-rescue factor A
+分子 #6: 50S ribosomal protein L1
+分子 #7: 50S ribosomal protein L2
+分子 #8: 50S ribosomal protein L3
+分子 #9: 50S ribosomal protein L4
+分子 #10: 50S ribosomal protein L5
+分子 #11: 50S ribosomal protein L6
+分子 #12: 50S ribosomal protein L9
+分子 #13: 50S ribosomal protein L10
+分子 #14: 50S ribosomal protein L11
+分子 #15: 50S ribosomal protein L13
+分子 #16: 50S ribosomal protein L14
+分子 #17: 50S ribosomal protein L15
+分子 #18: 50S ribosomal protein L16
+分子 #19: 50S ribosomal protein L17
+分子 #20: 50S ribosomal protein L18
+分子 #21: 50S ribosomal protein L19
+分子 #22: 50S ribosomal protein L20
+分子 #23: 50S ribosomal protein L21
+分子 #24: 50S ribosomal protein L22
+分子 #25: 50S ribosomal protein L23
+分子 #26: 50S ribosomal protein L24
+分子 #27: 50S ribosomal protein L25
+分子 #28: 50S ribosomal protein L27
+分子 #29: 50S ribosomal protein L28
+分子 #30: 50S ribosomal protein L29
+分子 #31: 50S ribosomal protein L30
+分子 #32: 50S ribosomal protein L31
+分子 #33: 50S ribosomal protein L32
+分子 #34: 50S ribosomal protein L33
+分子 #35: 50S ribosomal protein L34
+分子 #36: 50S ribosomal protein L35
+分子 #37: 50S ribosomal protein L36
+分子 #39: Peptide chain release factor RF2
+分子 #40: 30S ribosomal protein S2
+分子 #41: 30S ribosomal protein S3
+分子 #42: 30S ribosomal protein S4
+分子 #43: 30S ribosomal protein S5
+分子 #44: 30S ribosomal protein S6
+分子 #45: 30S ribosomal protein S7
+分子 #46: 30S ribosomal protein S8
+分子 #47: 30S ribosomal protein S9
+分子 #48: 30S ribosomal protein S10
+分子 #49: 30S ribosomal protein S11
+分子 #50: 30S ribosomal protein S12
+分子 #51: 30S ribosomal protein S13
+分子 #52: 30S ribosomal protein S14
+分子 #53: 30S ribosomal protein S15
+分子 #54: 30S ribosomal protein S16
+分子 #55: 30S ribosomal protein S17
+分子 #56: 30S ribosomal protein S18
+分子 #57: 30S ribosomal protein S19
+分子 #58: 30S ribosomal protein S20
+分子 #59: 30S ribosomal protein S21
+分子 #60: MAGNESIUM ION
+分子 #61: ZINC ION
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
緩衝液 | pH: 7 構成要素:
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グリッド | モデル: C-flat-2/2 / 材質: COPPER / メッシュ: 200 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: HOLEY / 前処理 - タイプ: PLASMA CLEANING / 前処理 - 時間: 20 sec. / 前処理 - 雰囲気: OTHER 詳細: Using a Solarus 950 plasma cleaning system. The forward RF target was set to 7w. | ||||||||||||||||||
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 95 % / チャンバー内温度: 277.15 K / 装置: FEI VITROBOT MARK IV |
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電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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撮影 | フィルム・検出器のモデル: DIRECT ELECTRON DE-20 (5k x 3k) 検出モード: COUNTING / デジタル化 - 画像ごとのフレーム数: 2-27 / 撮影したグリッド数: 1 / 実像数: 3760 / 平均電子線量: 1.2 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm / 最大 デフォーカス(公称値): 3.0 µm / 最小 デフォーカス(公称値): 0.5 µm / 倍率(公称値): 29000 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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画像解析
-原子モデル構築 1
精密化 | 空間: REAL / プロトコル: OTHER / 温度因子: 100 / 当てはまり具合の基準: Correlation coefficient |
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得られたモデル | ![]() PDB-5u9g: |