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基本情報
| 登録情報 | データベース: PDB / ID: 9ehz | ||||||||||||||||||||||||||||||
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| タイトル | Cryo-EM structure of Human RNA polymerase II Elongation Complex in an Intermediate Translocation State | ||||||||||||||||||||||||||||||
要素 |
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キーワード | TRANSCRIPTION / TRANSFERASE/DNA/RNA / translocation / Human RNA polymerase II / TRANSFERASE-DNA-RNA complex | ||||||||||||||||||||||||||||||
| 機能・相同性 | 機能・相同性情報microfibril binding / RNA Polymerase III Chain Elongation / RNA Polymerase III Transcription Termination / regulation of transcription by RNA polymerase I / 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 / RNA Polymerase III Abortive And Retractive Initiation / Cytosolic sensors of pathogen-associated DNA ...microfibril binding / RNA Polymerase III Chain Elongation / RNA Polymerase III Transcription Termination / regulation of transcription by RNA polymerase I / 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 / RNA Polymerase III Abortive And Retractive Initiation / Cytosolic sensors of pathogen-associated DNA / Abortive elongation of HIV-1 transcript in the absence of Tat / FGFR2 alternative splicing / RNA Polymerase I Transcription Termination / MicroRNA (miRNA) biogenesis / Viral Messenger RNA Synthesis / Signaling by FGFR2 IIIa TM / LRR domain binding / 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 / 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 / Formation of the Early Elongation Complex / Formation of the HIV-1 Early Elongation Complex / mRNA Capping / mRNA Splicing - Minor Pathway / PIWI-interacting RNA (piRNA) biogenesis / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / nuclear-transcribed mRNA catabolic process / Processing of Capped Intron-Containing Pre-mRNA / RNA Polymerase I Transcription Initiation / transcription by RNA polymerase III / positive regulation of translational initiation / Pausing and recovery of Tat-mediated HIV elongation / Tat-mediated HIV elongation arrest and recovery / 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 / Formation of HIV-1 elongation complex containing HIV-1 Tat / RNA polymerase I complex / transcription elongation by RNA polymerase I / RNA polymerase III complex / Formation of HIV elongation complex in the absence of HIV Tat / RNA polymerase II, core complex / tRNA transcription by RNA polymerase III / transcription by RNA polymerase I / RNA Polymerase II Transcription Elongation / Formation of RNA Pol II elongation complex / translation initiation factor binding / transcription-coupled nucleotide-excision repair / RNA Polymerase II Pre-transcription Events / mRNA Splicing - Major Pathway / Inhibition of DNA recombination at telomere / positive regulation of RNA splicing / TP53 Regulates Transcription of DNA Repair Genes / transcription initiation at RNA polymerase II promoter / promoter-specific chromatin binding / RNA Polymerase I Promoter Escape / P-body / Transcriptional regulation by small RNAs / DNA-templated transcription termination / protein-DNA complex / NoRC negatively regulates rRNA expression / B-WICH complex positively regulates rRNA expression / Transcription-Coupled Nucleotide Excision Repair (TC-NER) / Formation of TC-NER Pre-Incision Complex / kinase binding / ribonucleoside binding / Activation of anterior HOX genes in hindbrain development during early embryogenesis / fibrillar center / DNA-directed RNA polymerase / Dual incision in TC-NER / Gap-filling DNA repair synthesis and ligation in TC-NER / DNA-directed RNA polymerase activity / single-stranded DNA binding / chromosome / 加水分解酵素; エステル加水分解酵素; 5'-リン酸モノエステル産生エキソリボヌクレアーゼ / Estrogen-dependent gene expression / nucleic acid binding / transcription by RNA polymerase II / hydrolase activity / single-stranded RNA binding / protein dimerization activity / protein stabilization / nuclear speck / RNA-directed RNA polymerase / nucleotide binding / RNA-directed RNA polymerase activity / ubiquitin protein ligase binding / chromatin binding / regulation of DNA-templated transcription / nucleolus / magnesium ion binding / mitochondrion 類似検索 - 分子機能 | ||||||||||||||||||||||||||||||
| 生物種 | Homo sapiens (ヒト) | ||||||||||||||||||||||||||||||
| 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: 2.6 Å | ||||||||||||||||||||||||||||||
データ登録者 | Florez Ariza, A. / Lue, N. / Nogales, E. | ||||||||||||||||||||||||||||||
| 資金援助 | 米国, 2件
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引用 | ジャーナル: Nat Struct Mol Biol / 年: 2025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 poses a major challenge for genome stability. The RECQL5 helicase helps safeguard genome integrity and is the only member of the human RecQ helicase family that directly binds to RNA ...Transcription poses a major challenge for genome stability. The RECQL5 helicase helps safeguard genome integrity and is the only member of the human RecQ helicase family that directly binds to RNA polymerase II (Pol II) and affects its progression. While RECQL5 mitigates transcription stress in cells, the molecular mechanism underlying this phenomenon is unclear. Here, we use cryo-electron microscopy to determine the structures of stalled human 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, in its nucleotide-free state, RECQL5 twists the downstream DNA in the EC and, upon nucleotide binding, undergoes a conformational change that allosterically induces Pol II toward a post-translocation state. We propose that this mechanism may help restart Pol II elongation and, therefore, contribute to reducing transcription stress. #1: ジャーナル: bioRxiv / 年: 2025 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. | ||||||||||||||||||||||||||||||
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構造の表示
| 構造ビューア | 分子: Molmil Jmol/JSmol |
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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 | 9ehz.cif.gz | 823.7 KB | 表示 | PDBx/mmCIF形式 |
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| PDB形式 | pdb9ehz.ent.gz | 646.4 KB | 表示 | PDB形式 |
| PDBx/mmJSON形式 | 9ehz.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 | 9ehz_validation.pdf.gz | 1.3 MB | 表示 | wwPDB検証レポート |
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| 文書・詳細版 | 9ehz_full_validation.pdf.gz | 1.4 MB | 表示 | |
| XML形式データ | 9ehz_validation.xml.gz | 115.6 KB | 表示 | |
| CIF形式データ | 9ehz_validation.cif.gz | 179.7 KB | 表示 | |
| アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/eh/9ehz ftp://data.pdbj.org/pub/pdb/validation_reports/eh/9ehz | HTTPS FTP |
-関連構造データ
| 関連構造データ | ![]() 48071MC ![]() 9ei1C ![]() 9ei2C ![]() 9ei3C ![]() 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 F&H 検索 |
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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 DGIKABC
| #1: 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) Homo sapiens (ヒト) / 参照: UniProt: O15514 |
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| #4: 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) Homo sapiens (ヒト) / 参照: UniProt: P62487 |
| #6: 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) Homo sapiens (ヒト) / 参照: UniProt: P36954 |
| #8: 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) Homo sapiens (ヒト) / 参照: UniProt: P52435 |
| #13: 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) Homo sapiens (ヒト) / 参照: UniProt: P24928, DNA-directed RNA polymerase |
| #14: 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) Homo sapiens (ヒト) / 参照: UniProt: P30876, DNA-directed RNA polymerase |
| #15: 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) Homo sapiens (ヒト) / 参照: UniProt: P19387 |
-DNA-directed RNA polymerases I, II, and III subunit ... , 5種, 5分子 EFHJL
| #2: 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) Homo sapiens (ヒト) / 参照: UniProt: P19388 |
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| #3: 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) Homo sapiens (ヒト) / 参照: UniProt: P61218 |
| #5: 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) Homo sapiens (ヒト) / 参照: UniProt: P52434 |
| #7: 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) Homo sapiens (ヒト) / 参照: UniProt: P62875 |
| #9: 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) Homo sapiens (ヒト) / 参照: UniProt: P53803 |
-DNA鎖 , 2種, 2分子 NT
| #10: DNA鎖 | 分子量: 13305.593 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) Homo sapiens (ヒト) |
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| #12: DNA鎖 | 分子量: 8515.503 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) Homo sapiens (ヒト) |
-RNA鎖 , 1種, 1分子 P
| #11: RNA鎖 | 分子量: 6414.902 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) Homo sapiens (ヒト) |
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-非ポリマー , 2種, 9分子 


| #16: 化合物 | ChemComp-ZN / #17: 化合物 | ChemComp-MG / | |
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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 | N |
-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: Free Stalled Human RNA Polymerase II Elongation Complex in an Intermediate Translocation State タイプ: COMPLEX / Entity ID: #1-#15 / 由来: MULTIPLE SOURCES |
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| 分子量 | 値: 0.43 MDa / 実験値: NO |
| 由来(天然) | 生物種: Homo sapiens (ヒト) |
| 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: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: OTHER |
| 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: 2.6 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 174428 / 対称性のタイプ: POINT |
| 精密化 | 最高解像度: 2.6 Å 立体化学のターゲット値: REAL-SPACE (WEIGHTED MAP SUM AT ATOM CENTERS) |
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万見について




Homo sapiens (ヒト)
米国, 2件
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FIELD EMISSION GUN