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データを開く
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基本情報
登録情報 | データベース: PDB / ID: 8xrm | |||||||||||||||||||||||||||||||||
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タイトル | RNA polymerase II elongation complex with DSIF, SPT6, and ELOF1 transcribing genomic DNA extracted from human nuclei | |||||||||||||||||||||||||||||||||
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![]() | TRANSCRIPTION/RNA/DNA / nucleus / TRANSCRIPTION-RNA-DNA COMPLEX | |||||||||||||||||||||||||||||||||
機能・相同性 | ![]() microfibril binding / regulation of isotype switching / negative regulation of DNA-templated transcription, elongation / regulation of muscle cell differentiation / RNA Polymerase III Chain Elongation / RNA Polymerase III Transcription Termination / DSIF complex / regulation of transcription by RNA polymerase I / regulation of transcription elongation by RNA polymerase II / RPAP3/R2TP/prefoldin-like complex ...microfibril binding / regulation of isotype switching / negative regulation of DNA-templated transcription, elongation / regulation of muscle cell differentiation / RNA Polymerase III Chain Elongation / RNA Polymerase III Transcription Termination / DSIF complex / regulation of transcription by RNA polymerase I / regulation of transcription elongation by RNA polymerase II / RPAP3/R2TP/prefoldin-like complex / nucleosome organization / 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 / blastocyst formation / RNA Polymerase III Abortive And Retractive Initiation / positive regulation of DNA-templated transcription, elongation / Abortive elongation of HIV-1 transcript in the absence of Tat / RNA Polymerase I Transcription Termination / FGFR2 alternative splicing / MicroRNA (miRNA) biogenesis / Viral Messenger RNA Synthesis / LRR domain binding / Signaling by FGFR2 IIIa TM / transcription elongation-coupled chromatin remodeling / RNA Pol II CTD phosphorylation and interaction with CE during HIV infection / RNA Pol II CTD phosphorylation and interaction with CE / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / Formation of the Early Elongation Complex / Formation of the HIV-1 Early Elongation Complex / 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 Capping / mRNA Splicing - Minor Pathway / PIWI-interacting RNA (piRNA) biogenesis / RNA polymerase II complex binding / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / RNA Polymerase I Transcription Initiation / nuclear-transcribed mRNA catabolic process / Processing of Capped Intron-Containing Pre-mRNA / Pausing and recovery of Tat-mediated HIV elongation / Tat-mediated HIV elongation arrest and recovery / positive regulation of translational initiation / negative regulation of transcription elongation by RNA polymerase II / positive regulation of macroautophagy / transcription by RNA polymerase III / RNA polymerase II transcribes snRNA genes / HIV elongation arrest and recovery / Pausing and recovery of HIV elongation / mRNA transport / Tat-mediated elongation of the HIV-1 transcript / Formation of HIV-1 elongation complex containing HIV-1 Tat / RNA polymerase I complex / Formation of HIV elongation complex in the absence of HIV Tat / transcription elongation by RNA polymerase I / RNA polymerase III complex / RNA polymerase II, core complex / nucleosome binding / tRNA transcription by RNA polymerase III / RNA Polymerase II Transcription Elongation / transcription by RNA polymerase I / Formation of RNA Pol II elongation complex / translation initiation factor binding / transcription-coupled nucleotide-excision repair / RNA Polymerase II Pre-transcription Events / Inhibition of DNA recombination at telomere / mRNA Splicing - Major Pathway / RNA splicing / positive regulation of RNA splicing / transcription elongation factor complex / TP53 Regulates Transcription of DNA Repair Genes / transcription initiation at RNA polymerase II promoter / RNA Polymerase I Promoter Escape / transcription elongation by RNA polymerase II / P-body / promoter-specific chromatin binding / Transcriptional regulation by small RNAs / Transcription-Coupled Nucleotide Excision Repair (TC-NER) / positive regulation of transcription elongation by RNA polymerase II / Formation of TC-NER Pre-Incision Complex / DNA-templated transcription termination / protein-DNA complex / NoRC negatively regulates rRNA expression / B-WICH complex positively regulates rRNA expression / Dual incision in TC-NER / Gap-filling DNA repair synthesis and ligation in TC-NER / ribonucleoside binding / kinase binding / Activation of anterior HOX genes in hindbrain development during early embryogenesis / fibrillar center / mRNA processing / DNA-directed RNA polymerase / DNA-directed RNA polymerase activity / single-stranded DNA binding 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: 3.13 Å | |||||||||||||||||||||||||||||||||
![]() | Kujirai, T. / Kato, J. / Yamamoto, K. / Hirai, S. / Negishi, L. / Ogasawara, M. / Takizawa, Y. / Kurumizaka, H. | |||||||||||||||||||||||||||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Multiple structures of RNA polymerase II isolated from human nuclei by ChIP-CryoEM analysis. 著者: Tomoya Kujirai / Junko Kato / Kyoka Yamamoto / Seiya Hirai / Takeru Fujii / Kazumitsu Maehara / Akihito Harada / Lumi Negishi / Mitsuo Ogasawara / Yuki Yamaguchi / Yasuyuki Ohkawa / Yoshimasa ...著者: Tomoya Kujirai / Junko Kato / Kyoka Yamamoto / Seiya Hirai / Takeru Fujii / Kazumitsu Maehara / Akihito Harada / Lumi Negishi / Mitsuo Ogasawara / Yuki Yamaguchi / Yasuyuki Ohkawa / Yoshimasa Takizawa / Hitoshi Kurumizaka / ![]() 要旨: RNA polymerase II (RNAPII) is a central transcription enzyme that exists as multiple forms with or without accessory factors, and transcribes the genomic DNA packaged in chromatin. To understand how ...RNA polymerase II (RNAPII) is a central transcription enzyme that exists as multiple forms with or without accessory factors, and transcribes the genomic DNA packaged in chromatin. To understand how RNAPII functions in the human genome, we isolate transcribing RNAPII complexes from human nuclei by chromatin immunopurification, and determine the cryo-electron microscopy structures of RNAPII elongation complexes (ECs) associated with genomic DNA in distinct forms, without or with the elongation factors SPT4/5, ELOF1, and SPT6. This ChIP-cryoEM method also reveals the two EC-nucleosome complexes corresponding nucleosome disassembly/reassembly processes. In the structure of EC-downstream nucleosome, EC paused at superhelical location (SHL) -5 in the nucleosome, suggesting that SHL(-5) pausing occurs in a sequence-independent manner during nucleosome disassembly. In the structure of the EC-upstream nucleosome, EC directly contacts the nucleosome through the nucleosomal DNA-RPB4/7 stalk and the H2A-H2B dimer-RPB2 wall interactions, suggesting that EC may be paused during nucleosome reassembly. These representative EC structures transcribing the human genome provide mechanistic insights into understanding RNAPII transcription on chromatin. | |||||||||||||||||||||||||||||||||
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 | ![]() | 816 KB | 表示 | ![]() |
PDBx/mmJSON形式 | ![]() | ツリー表示 | ![]() | |
その他 | ![]() |
-検証レポート
文書・要旨 | ![]() | 1.3 MB | 表示 | ![]() |
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文書・詳細版 | ![]() | 1.3 MB | 表示 | |
XML形式データ | ![]() | 137.5 KB | 表示 | |
CIF形式データ | ![]() | 221.9 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
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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) ![]() 参照: UniProt: P24928, DNA-directed RNA polymerase, RNA-directed RNA polymerase |
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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: 33630.387 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) ![]() ![]() |
#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: 24584.223 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) ![]() |
-Transcription elongation factor ... , 4種, 4分子 MQYZ
#13: タンパク質 | 分子量: 199330.719 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
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#16: タンパク質 | 分子量: 9475.881 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
#18: タンパク質 | 分子量: 13210.201 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
#19: タンパク質 | 分子量: 121145.477 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) ![]() |
-DNA鎖 , 2種, 2分子 NT
#14: DNA鎖 | 分子量: 13708.779 Da / 分子数: 1 / 由来タイプ: 天然 詳細: The DNA sequence in this structure is unknown because it is an average of genomic DNA regions bound to the RNA polymerase II elongation complex. 由来: (天然) ![]() |
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#17: DNA鎖 | 分子量: 13396.525 Da / 分子数: 1 / 由来タイプ: 天然 詳細: The DNA sequence in this structure is unknown because it is an average of genomic DNA regions bound to the RNA polymerase II elongation complex. 由来: (天然) ![]() |
-RNA鎖 , 1種, 1分子 P
#15: RNA鎖 | 分子量: 6773.156 Da / 分子数: 1 / 由来タイプ: 天然 詳細: The RNA sequence in this structure is unknown because it is an average of RNA bound to the RNA polymerase II elongation complex on genomic DNA. 由来: (天然) ![]() |
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-非ポリマー , 2種, 11分子 


#20: 化合物 | ChemComp-ZN / #21: 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 | N |
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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: RNA polymerase II complexes isolated from HeLa cell nuclei タイプ: COMPLEX / Entity ID: #1-#19 / 由来: NATURAL |
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由来(天然) | 生物種: ![]() |
由来(組換発現) | 生物種: ![]() |
緩衝液 | pH: 7.5 |
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): 2500 nm / 最小 デフォーカス(公称値): 1000 nm |
撮影 | 電子線照射量: 60 e/Å2 フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) |
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解析
EMソフトウェア | 名称: PHENIX / バージョン: 1.19.2_4158: / 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: 3.13 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 132687 / 対称性のタイプ: POINT | ||||||||||||||||||||||||
拘束条件 |
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