+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-6667 | |||||||||
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タイトル | Mechanistic insights into the alternative translation termination by ArfA and RF2 | |||||||||
マップデータ | ||||||||||
試料 |
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キーワード | ribosome / translation / RF2 / Arfa / non-stop mRNA / ribosome rescue | |||||||||
機能・相同性 | 機能・相同性情報 translation release factor activity, codon specific / 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 / endoribonuclease inhibitor activity ...translation release factor activity, codon specific / 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 / 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 / rescue of stalled ribosome / 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 / 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 / rRNA binding / ribosome / structural constituent of ribosome / translation / viral translational frameshifting / 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) / Escherichia coli K-12 (大腸菌) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.01 Å | |||||||||
データ登録者 | Ma C / Kurita D | |||||||||
引用 | ジャーナル: Nature / 年: 2017 タイトル: Mechanistic insights into the alternative translation termination by ArfA and RF2. 著者: Chengying Ma / Daisuke Kurita / Ningning Li / Yan Chen / Hyouta Himeno / Ning Gao / 要旨: During cellular translation of messenger RNAs by ribosomes, the translation apparatus sometimes pauses or stalls at the elongation and termination steps. With the exception of programmed stalling, ...During cellular translation of messenger RNAs by ribosomes, the translation apparatus sometimes pauses or stalls at the elongation and termination steps. With the exception of programmed stalling, which is usually used by cells for regulatory purposes, ribosomes stalled on mRNAs need to be terminated and recycled to maintain adequate translation capacity. Much ribosome stalling originates in aberrant mRNAs that lack a stop codon. Transcriptional errors, misprocessing of primary transcripts, and undesired mRNA cleavage all contribute to the formation of non-stop mRNAs. Ribosomes stalled at the 3' end of non-stop mRNAs do not undergo normal termination owing to the lack of specific stop-codon recognition by canonical peptide release factors at the A-site decoding centre. In bacteria, the transfer-messenger RNA (tmRNA)-SmpB-mediated trans-translation rescue system reroutes stalled ribosomes to the normal elongation cycle and translation termination. Two additional rescue systems, ArfA-RF2 (refs 13, 14, 15, 16) and ArfB (formerly known as YaeJ), are also present in many bacterial species, but their mechanisms are not fully understood. Here, using cryo-electron microscopy, we characterize the structure of the Escherichia coli 70S ribosome bound with ArfA, the release factor RF2, a short non-stop mRNA and a cognate P-site tRNA. The C-terminal loop of ArfA occupies the mRNA entry channel on the 30S subunit, whereas its N terminus is sandwiched between the decoding centre and the switch loop of RF2, leading to marked conformational changes in both the decoding centre and RF2. Despite the distinct conformation of RF2, its conserved catalytic GGQ motif is precisely positioned next to the CCA-end of the P-site tRNA. These data illustrate a stop-codon surrogate mechanism for ArfA in facilitating the termination of non-stop ribosomal complexes by RF2. | |||||||||
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 | EMマップ: SurfViewMolmilJmol/JSmol |
添付画像 |
-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_6667.map.gz | 16.4 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-6667-v30.xml emd-6667.xml | 77.6 KB 77.6 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_6667.png | 72.4 KB | ||
Filedesc metadata | emd-6667.cif.gz | 13.3 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-6667 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-6667 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_6667_validation.pdf.gz | 478.6 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_6667_full_validation.pdf.gz | 478.2 KB | 表示 | |
XML形式データ | emd_6667_validation.xml.gz | 7 KB | 表示 | |
CIF形式データ | emd_6667_validation.cif.gz | 8 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-6667 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-6667 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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「今月の分子」の関連する項目 |
-マップ
ファイル | ダウンロード / ファイル: emd_6667.map.gz / 形式: CCP4 / 大きさ: 178 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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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: 1.08 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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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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-添付データ
-試料の構成要素
+全体 : 70S-ArfA-RF2 complex
+超分子 #1: 70S-ArfA-RF2 complex
+超分子 #2: Rf2-ArfA
+超分子 #3: rRNA
+分子 #1: 30S ribosomal protein S20
+分子 #2: 30S ribosomal protein S21
+分子 #3: Alternative ribosome-rescue factor A
+分子 #4: Peptide chain release factor 2
+分子 #9: 50S ribosomal protein L2
+分子 #10: 50S ribosomal protein L3
+分子 #11: 50S ribosomal protein L4
+分子 #12: 50S ribosomal protein L5
+分子 #13: 50S ribosomal protein L6
+分子 #14: 50S ribosomal protein L9
+分子 #15: 50S ribosomal protein L10
+分子 #16: 50S ribosomal protein L11
+分子 #17: 50S ribosomal protein L13
+分子 #18: 50S ribosomal protein L14
+分子 #19: 50S ribosomal protein L15
+分子 #20: 50S ribosomal protein L16
+分子 #21: 50S ribosomal protein L17
+分子 #22: 50S ribosomal protein L18
+分子 #23: 50S ribosomal protein L19
+分子 #24: 50S ribosomal protein L20
+分子 #25: 50S ribosomal protein L21
+分子 #26: 50S ribosomal protein L22
+分子 #27: 50S ribosomal protein L23
+分子 #28: 50S ribosomal protein L24
+分子 #29: 50S ribosomal protein L25
+分子 #30: 50S ribosomal protein L27
+分子 #31: 50S ribosomal protein L28
+分子 #32: 50S ribosomal protein L29
+分子 #33: 50S ribosomal protein L30
+分子 #34: 50S ribosomal protein L32
+分子 #35: 50S ribosomal protein L33
+分子 #36: 50S ribosomal protein L34
+分子 #37: 50S ribosomal protein L35
+分子 #38: 50S ribosomal protein L36
+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 #5: P-site tRNA
+分子 #6: mRNA
+分子 #7: 23S rRNA
+分子 #8: 5S Ribosomal RNA
+分子 #39: 16S rRNA
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
緩衝液 | pH: 7.6 |
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グリッド | 材質: COPPER / 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: GLOW DISCHARGE / 前処理 - 時間: 30 sec. |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277.15 K |
-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: FEI FALCON II (4k x 4k) 検出モード: INTEGRATING / 平均電子線量: 2.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD |
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 |
-画像解析
初期モデル | モデルのタイプ: OTHER |
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最終 再構成 | 使用したクラス数: 8 / 解像度のタイプ: BY AUTHOR / 解像度: 3.01 Å / 解像度の算出法: FSC 0.143 CUT-OFF / ソフトウェア - 名称: RELION (ver. 1.3) / 使用した粒子像数: 13693 |
初期 角度割当 | タイプ: PROJECTION MATCHING |
最終 角度割当 | タイプ: PROJECTION MATCHING |