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基本情報
登録情報 | データベース: PDB / ID: 9ei3 | ||||||||||||||||||||||||||||||
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タイトル | Cryo-EM structure of Human RNA polymerase II Elongation Complex bound to the RECQL5 helicase in the presence of AMPPNP | ||||||||||||||||||||||||||||||
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![]() | TRANSCRIPTION / TRANSFERASE/DNA/RNA / translocation / Human RNA polymerase II / RECQL5 helicase / TRANSFERASE-DNA-RNA complex | ||||||||||||||||||||||||||||||
機能・相同性 | ![]() mitotic DNA-templated DNA replication / LRR domain binding / microfibril binding / chromosome separation / cellular response to camptothecin / RNA Polymerase III Chain Elongation / RNA Polymerase III Transcription Termination / regulation of transcription by RNA polymerase I / replication-born double-strand break repair via sister chromatid exchange / RPAP3/R2TP/prefoldin-like complex ...mitotic DNA-templated DNA replication / LRR domain binding / microfibril binding / chromosome separation / cellular response to camptothecin / RNA Polymerase III Chain Elongation / RNA Polymerase III Transcription Termination / regulation of transcription by RNA polymerase I / replication-born double-strand break repair via sister chromatid exchange / RPAP3/R2TP/prefoldin-like complex / RNA Polymerase III Transcription Initiation From Type 1 Promoter / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA Polymerase III Transcription Initiation From Type 3 Promoter / Cytosolic sensors of pathogen-associated DNA / RNA Polymerase III Abortive And Retractive Initiation / Abortive elongation of HIV-1 transcript in the absence of Tat / FGFR2 alternative splicing / RNA Polymerase I Transcription Termination / transcription preinitiation complex / MicroRNA (miRNA) biogenesis / 3'-5' DNA helicase activity / DNA 3'-5' helicase / Viral Messenger RNA Synthesis / Signaling by FGFR2 IIIa TM / DNA metabolic process / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / RNA Pol II CTD phosphorylation and interaction with CE during HIV infection / RNA Pol II CTD phosphorylation and interaction with CE / Formation of the Early Elongation Complex / Formation of the HIV-1 Early Elongation Complex / mRNA Capping / HIV Transcription Initiation / RNA Polymerase II HIV Promoter Escape / Transcription of the HIV genome / RNA Polymerase II Promoter Escape / RNA Polymerase II Transcription Pre-Initiation And Promoter Opening / RNA Polymerase II Transcription Initiation / RNA Polymerase II Transcription Initiation And Promoter Clearance / mRNA Splicing - Minor Pathway / RNA polymerase II complex binding / PIWI-interacting RNA (piRNA) biogenesis / negative regulation of transcription elongation by RNA polymerase II / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / RNA Polymerase I Transcription Initiation / nuclear-transcribed mRNA catabolic process / Pausing and recovery of Tat-mediated HIV elongation / Tat-mediated HIV elongation arrest and recovery / Processing of Capped Intron-Containing Pre-mRNA / positive regulation of translational initiation / RNA polymerase II transcribes snRNA genes / HIV elongation arrest and recovery / Pausing and recovery of HIV elongation / Tat-mediated elongation of the HIV-1 transcript / negative regulation of double-strand break repair via homologous recombination / Formation of HIV-1 elongation complex containing HIV-1 Tat / transcription by RNA polymerase III / transcription by RNA polymerase I / RNA polymerase I complex / transcription elongation by RNA polymerase I / Formation of HIV elongation complex in the absence of HIV Tat / RNA polymerase III complex / transcription-coupled nucleotide-excision repair / DNA helicase activity / RNA polymerase II, core complex / tRNA transcription by RNA polymerase III / RNA Polymerase II Transcription Elongation / : / Formation of RNA Pol II elongation complex / translation initiation factor binding / RNA Polymerase II Pre-transcription Events / DNA-directed RNA polymerase activity / Inhibition of DNA recombination at telomere / mRNA Splicing - Major Pathway / positive regulation of RNA splicing / replication fork / isomerase activity / helicase activity / TP53 Regulates Transcription of DNA Repair Genes / transcription initiation at RNA polymerase II promoter / RNA Polymerase I Promoter Escape / Transcriptional regulation by small RNAs / promoter-specific chromatin binding / P-body / DNA-templated transcription termination / double-strand break repair via homologous recombination / protein-DNA complex / NoRC negatively regulates rRNA expression / B-WICH complex positively regulates rRNA expression / Transcription-Coupled Nucleotide Excision Repair (TC-NER) / ribonucleoside binding / Formation of TC-NER Pre-Incision Complex / kinase binding / Activation of anterior HOX genes in hindbrain development during early embryogenesis / : / : / : / fibrillar center / : / : / : 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: 3.2 Å | ||||||||||||||||||||||||||||||
![]() | Florez Ariza, A. / Lue, N. / Nogales, E. | ||||||||||||||||||||||||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Structural insights into transcriptional regulation by the helicase RECQL5. 著者: Alfredo Jose Florez Ariza / Nicholas Z Lue / Patricia Grob / Benjamin Kaeser / Jie Fang / Susanne A Kassube / Eva Nogales / ![]() 要旨: Transcription and its regulation pose a major challenge for genome stability. The helicase RECQL5 has been proposed as an important factor to help safeguard the genome, and is the only member of the ...Transcription and its regulation pose a major challenge for genome stability. The helicase RECQL5 has been proposed as an important factor to help safeguard the genome, and is the only member of the human RecQ helicase family that directly binds to RNA Polymerase II (Pol II) and affects its progression. RECQL5 mitigates transcription stress and genome instability in cells, yet the molecular mechanism underlying this phenomenon is unclear. Here, we employ cryo-electron microscopy (cryo-EM) to determine the structures of stalled Pol II elongation complexes (ECs) bound to RECQL5. Our structures reveal the molecular interactions stabilizing RECQL5 binding to the Pol II EC and highlight its role as a transcriptional roadblock. Additionally, we find that RECQL5 can modulate the Pol II translocation state. In its nucleotide-free state, RECQL5 mechanically twists the downstream DNA in the EC, and upon nucleotide binding, it undergoes a conformational change that allosterically induces Pol II towards a post-translocation state. We propose this mechanism may help restart Pol II elongation and therefore contribute to reduction of transcription stress. | ||||||||||||||||||||||||||||||
履歴 |
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構造の表示
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 | ![]() | 1 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
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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-関連構造データ
関連構造データ | ![]() 48075MC ![]() 9ehzC ![]() 9ei1C ![]() 9ei2C ![]() 9ei4C M: このデータのモデリングに利用したマップデータ C: 同じ文献を引用 ( |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
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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
-DNA-directed RNA polymerase II subunit ... , 7種, 7分子 ABCDGIK
#1: タンパク質 | 分子量: 217420.047 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
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#2: タンパク質 | 分子量: 134071.453 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
#3: タンパク質 | 分子量: 31478.148 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
#4: タンパク質 | 分子量: 16331.255 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
#7: タンパク質 | 分子量: 19314.283 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
#9: タンパク質 | 分子量: 14541.221 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
#11: タンパク質 | 分子量: 13310.284 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
-DNA-directed RNA polymerases I, II, and III subunit ... , 5種, 5分子 EFHJL
#5: タンパク質 | 分子量: 24644.318 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
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#6: タンパク質 | 分子量: 14491.026 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
#8: タンパク質 | 分子量: 17162.273 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
#10: タンパク質 | 分子量: 7655.123 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
#12: タンパク質 | 分子量: 7018.244 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
-DNA鎖 , 2種, 2分子 NT
#13: DNA鎖 | 分子量: 13305.593 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) ![]() |
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#15: DNA鎖 | 分子量: 8515.503 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) ![]() |
-RNA鎖 / タンパク質 , 2種, 2分子 PU
#14: RNA鎖 | 分子量: 6414.902 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) ![]() |
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#16: タンパク質 | 分子量: 109024.859 Da / 分子数: 1 / 由来タイプ: 組換発現 詳細: ATP-dependent DNA helicase Q5 bound to the nucleotide analog AMPPNP (ANP) 由来: (組換発現) ![]() ![]() ![]() |
-非ポリマー , 3種, 10分子 




#17: 化合物 | ChemComp-ZN / #18: 化合物 | ChemComp-MG / | #19: 化合物 | ChemComp-ANP / | |
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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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Has protein modification | Y |
-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 | 名称: Stalled Human RNA polymerase II Elongation Complex bound to an AMPPNP-bound RECQL5 helicase タイプ: COMPLEX / Entity ID: #1-#16 / 由来: MULTIPLE SOURCES |
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分子量 | 値: 0.49 MDa / 実験値: NO |
由来(天然) | 生物種: ![]() |
緩衝液 | pH: 8 |
試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
急速凍結 | 凍結剤: ETHANE |
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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: TFS KRIOS |
電子銃 | 電子線源: ![]() |
電子レンズ | モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 1800 nm / 最小 デフォーカス(公称値): 800 nm |
撮影 | 電子線照射量: 50 e/Å2 / フィルム・検出器のモデル: GATAN K3 (6k x 4k) |
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解析
EMソフトウェア | 名称: PHENIX / バージョン: 1.20.1_4487 / カテゴリ: モデル精密化 |
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CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION |
3次元再構成 | 解像度: 3.2 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 80622 / 対称性のタイプ: POINT |
精密化 | 交差検証法: NONE |