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基本情報
登録情報 | データベース: PDB / ID: 8xrj | |||||||||||||||||||||||||||||||||
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タイトル | RNA polymerase II elongation complex with upstream nucleosome extracted from human nuclei | |||||||||||||||||||||||||||||||||
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![]() | TRANSCRIPTION/DNA/RNA / nucleus / nucleosome / TRANSCRIPTION-DNA-RNA COMPLEX | |||||||||||||||||||||||||||||||||
機能・相同性 | ![]() LRR domain binding / 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 / Cytosolic sensors of pathogen-associated DNA ...LRR domain binding / 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 / 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 / MicroRNA (miRNA) biogenesis / Viral Messenger RNA Synthesis / Signaling by FGFR2 IIIa TM / 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 / PIWI-interacting RNA (piRNA) biogenesis / 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 / negative regulation of tumor necrosis factor-mediated signaling pathway / Tat-mediated elongation of the HIV-1 transcript / Formation of HIV-1 elongation complex containing HIV-1 Tat / transcription by RNA polymerase III / transcription by RNA polymerase I / negative regulation of megakaryocyte differentiation / protein localization to CENP-A containing chromatin / 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 / Chromatin modifying enzymes / Replacement of protamines by nucleosomes in the male pronucleus / RNA polymerase II, core complex / CENP-A containing nucleosome / tRNA transcription by RNA polymerase III / RNA Polymerase II Transcription Elongation / : / Packaging Of Telomere Ends / Formation of RNA Pol II elongation complex / Recognition and association of DNA glycosylase with site containing an affected purine / Cleavage of the damaged purine / translation initiation factor binding / Recognition and association of DNA glycosylase with site containing an affected pyrimidine / Cleavage of the damaged pyrimidine / RNA Polymerase II Pre-transcription Events / Deposition of new CENPA-containing nucleosomes at the centromere / telomere organization / DNA-directed RNA polymerase activity / Inhibition of DNA recombination at telomere / Meiotic synapsis / Interleukin-7 signaling / RNA Polymerase I Promoter Opening / Assembly of the ORC complex at the origin of replication / mRNA Splicing - Major Pathway / Regulation of endogenous retroelements by the Human Silencing Hub (HUSH) complex / innate immune response in mucosa / SUMOylation of chromatin organization proteins / DNA methylation / Condensation of Prophase Chromosomes / positive regulation of RNA splicing / Chromatin modifications during the maternal to zygotic transition (MZT) / HCMV Late Events / SIRT1 negatively regulates rRNA expression / epigenetic regulation of gene expression / ERCC6 (CSB) and EHMT2 (G9a) positively regulate rRNA expression / PRC2 methylates histones and DNA / Regulation of endogenous retroelements by KRAB-ZFP proteins / Defective pyroptosis / Regulation of endogenous retroelements by Piwi-interacting RNAs (piRNAs) / HDACs deacetylate histones / Nonhomologous End-Joining (NHEJ) / TP53 Regulates Transcription of DNA Repair Genes / transcription initiation at RNA polymerase II promoter / RNA Polymerase I Promoter Escape / lipopolysaccharide binding / Transcriptional regulation by small RNAs 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: 3.6 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 | ![]() | 985.4 KB | 表示 | ![]() |
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PDB形式 | ![]() | 773.9 KB | 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
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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) ![]() |
-DNA鎖 , 2種, 2分子 NT
#13: DNA鎖 | 分子量: 48979.957 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. 由来: (天然) ![]() |
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#15: DNA鎖 | 分子量: 48560.262 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. 由来: (天然) ![]() |
-RNA鎖 , 1種, 1分子 P
#14: RNA鎖 | 分子量: 3529.194 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 on genomic DNA. 由来: (天然) ![]() |
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-タンパク質 , 4種, 8分子 aebfcgdh
#16: タンパク質 | 分子量: 15305.969 Da / 分子数: 2 / 由来タイプ: 天然 / 由来: (天然) ![]() #17: タンパク質 | 分子量: 11263.231 Da / 分子数: 2 / 由来タイプ: 天然 / 由来: (天然) ![]() #18: タンパク質 | 分子量: 14034.355 Da / 分子数: 2 / 由来タイプ: 天然 / 由来: (天然) ![]() #19: タンパク質 | 分子量: 13804.045 Da / 分子数: 2 / 由来タイプ: 天然 / 由来: (天然) ![]() |
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-非ポリマー , 2種, 9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: 1.20.1_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: 3.6 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 32065 / 対称性のタイプ: POINT |