+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 8he5 | |||||||||||||||||||||||||||
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タイトル | RNA polymerase II elongation complex bound with Rad26 and Elf1, stalled at SHL(-3.5) of the nucleosome | |||||||||||||||||||||||||||
要素 |
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キーワード | TRANSCRIPTION / RNA / DNA / Repair | |||||||||||||||||||||||||||
機能・相同性 | 機能・相同性情報 regulation of septum digestion after cytokinesis / siRNA-mediated pericentric heterochromatin formation / RPB4-RPB7 complex / ATP-dependent chromatin remodeler activity / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / termination of RNA polymerase II transcription / termination of RNA polymerase III transcription / termination of RNA polymerase I transcription / RNA polymerase III activity / transcription initiation at RNA polymerase III promoter ...regulation of septum digestion after cytokinesis / siRNA-mediated pericentric heterochromatin formation / RPB4-RPB7 complex / ATP-dependent chromatin remodeler activity / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / termination of RNA polymerase II transcription / termination of RNA polymerase III transcription / termination of RNA polymerase I transcription / RNA polymerase III activity / transcription initiation at RNA polymerase III promoter / RNA polymerase II complex binding / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / protein localization to CENP-A containing chromatin / transcription initiation at RNA polymerase I promoter / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / CENP-A containing nucleosome / RNA polymerase II activity / negative regulation of tumor necrosis factor-mediated signaling pathway / Replacement of protamines by nucleosomes in the male pronucleus / transcription elongation by RNA polymerase I / arachidonate 15-lipoxygenase / arachidonate 15-lipoxygenase activity / tRNA transcription by RNA polymerase III / RNA polymerase I activity / Packaging Of Telomere Ends / RNA polymerase I complex / RNA polymerase III complex / positive regulation of translational initiation / transcription-coupled nucleotide-excision repair / lipoxygenase pathway / RNA polymerase II, core complex / Recognition and association of DNA glycosylase with site containing an affected purine / Cleavage of the damaged purine / pericentric heterochromatin / arachidonate metabolic process / lipid oxidation / Deposition of new CENPA-containing nucleosomes at the centromere / hepoxilin biosynthetic process / Recognition and association of DNA glycosylase with site containing an affected pyrimidine / Cleavage of the damaged pyrimidine / linoleic acid metabolic process / Meiotic synapsis / Inhibition of DNA recombination at telomere / nucleosomal DNA binding / translation initiation factor binding / RNA Polymerase I Promoter Opening / Assembly of the ORC complex at the origin of replication / DNA methylation / Condensation of Prophase Chromosomes / HCMV Late Events / SIRT1 negatively regulates rRNA expression / Chromatin modifications during the maternal to zygotic transition (MZT) / ERCC6 (CSB) and EHMT2 (G9a) positively regulate rRNA expression / PRC2 methylates histones and DNA / transcription elongation factor complex / Defective pyroptosis / Meiotic recombination / innate immune response in mucosa / DNA Damage/Telomere Stress Induced Senescence / HDACs deacetylate histones / Nonhomologous End-Joining (NHEJ) / RNA Polymerase I Promoter Escape / Transcriptional regulation by small RNAs / transcription initiation at RNA polymerase II promoter / lipopolysaccharide binding / Transcriptional regulation of granulopoiesis / transcription elongation by RNA polymerase II / Formation of the beta-catenin:TCF transactivating complex / HCMV Early Events / RUNX1 regulates genes involved in megakaryocyte differentiation and platelet function / P-body / G2/M DNA damage checkpoint / NoRC negatively regulates rRNA expression / Activated PKN1 stimulates transcription of AR (androgen receptor) regulated genes KLK2 and KLK3 / B-WICH complex positively regulates rRNA expression / heterochromatin formation / RMTs methylate histone arginines / ribonucleoside binding / Pre-NOTCH Transcription and Translation / Metalloprotease DUBs / Activation of anterior HOX genes in hindbrain development during early embryogenesis / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / structural constituent of chromatin / UCH proteinases / nucleosome / antimicrobial humoral immune response mediated by antimicrobial peptide / Processing of DNA double-strand break ends / nucleosome assembly / Recruitment and ATM-mediated phosphorylation of repair and signaling proteins at DNA double strand breaks / chromatin organization / E3 ubiquitin ligases ubiquitinate target proteins / Senescence-Associated Secretory Phenotype (SASP) / RUNX1 regulates transcription of genes involved in differentiation of HSCs / HATs acetylate histones / single-stranded DNA binding / antibacterial humoral response / Oxidative Stress Induced Senescence / defense response to Gram-negative bacterium / Estrogen-dependent gene expression 類似検索 - 分子機能 | |||||||||||||||||||||||||||
生物種 | Komagataella phaffii (菌類) Homo sapiens (ヒト) synthetic construct (人工物) | |||||||||||||||||||||||||||
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: 6.95 Å | |||||||||||||||||||||||||||
データ登録者 | Osumi, K. / Kujirai, T. / Ehara, H. / Kinoshita, C. / Saotome, M. / Kagawa, W. / Sekine, S. / Takizawa, Y. / Kurumizaka, H. | |||||||||||||||||||||||||||
資金援助 | 日本, 8件
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引用 | ジャーナル: J Mol Biol / 年: 2023 タイトル: Structural Basis of Damaged Nucleotide Recognition by Transcribing RNA Polymerase II in the Nucleosome. 著者: Ken Osumi / Tomoya Kujirai / Haruhiko Ehara / Mitsuo Ogasawara / Chiaki Kinoshita / Mika Saotome / Wataru Kagawa / Shun-Ichi Sekine / Yoshimasa Takizawa / Hitoshi Kurumizaka / 要旨: In transcription-coupled repair (TCR), transcribing RNA polymerase II (RNAPII) stalls at a DNA lesion and recruits TCR proteins to the damaged site. However, the mechanism by which RNAPII recognizes ...In transcription-coupled repair (TCR), transcribing RNA polymerase II (RNAPII) stalls at a DNA lesion and recruits TCR proteins to the damaged site. However, the mechanism by which RNAPII recognizes a DNA lesion in the nucleosome remains enigmatic. In the present study, we inserted an apurinic/apyrimidinic DNA lesion analogue, tetrahydrofuran (THF), in the nucleosomal DNA, where RNAPII stalls at the SHL(-4), SHL(-3.5), and SHL(-3) positions, and determined the structures of these complexes by cryo-electron microscopy. In the RNAPII-nucleosome complex stalled at SHL(-3.5), the nucleosome orientation relative to RNAPII is quite different from those in the SHL(-4) and SHL(-3) complexes, which have nucleosome orientations similar to naturally paused RNAPII-nucleosome complexes. Furthermore, we found that an essential TCR protein, Rad26 (CSB), enhances the RNAPII processivity, and consequently augments the DNA damage recognition efficiency of RNAPII in the nucleosome. The cryo-EM structure of the Rad26-RNAPII-nucleosome complex revealed that Rad26 binds to the stalled RNAPII through a novel interface, which is completely different from those previously reported. These structures may provide important information to understand the mechanism by which RNAPII recognizes the nucleosomal DNA lesion and recruits TCR proteins to the stalled RNAPII on the nucleosome. | |||||||||||||||||||||||||||
履歴 |
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-構造の表示
構造ビューア | 分子: MolmilJmol/JSmol |
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-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 8he5.cif.gz | 1.1 MB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb8he5.ent.gz | 872.9 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 8he5.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 | 8he5_validation.pdf.gz | 1.6 MB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 8he5_full_validation.pdf.gz | 1.6 MB | 表示 | |
XML形式データ | 8he5_validation.xml.gz | 134.1 KB | 表示 | |
CIF形式データ | 8he5_validation.cif.gz | 220.1 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/he/8he5 ftp://data.pdbj.org/pub/pdb/validation_reports/he/8he5 | HTTPS FTP |
-関連構造データ
-リンク
-集合体
登録構造単位 |
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-要素
-DNA-directed RNA polymerase ... , 3種, 3分子 ABI
#1: タンパク質 | 分子量: 194107.422 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Komagataella phaffii (菌類) / 参照: UniProt: C4R4Y0, DNA-directed RNA polymerase |
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#2: タンパク質 | 分子量: 139746.094 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Komagataella phaffii (菌類) / 参照: UniProt: C4QZQ7, DNA-directed RNA polymerase |
#9: タンパク質 | 分子量: 13612.320 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Komagataella phaffii (菌類) / 参照: UniProt: F2QPE6 |
-RNA polymerase II ... , 4種, 4分子 CDGK
#3: タンパク質 | 分子量: 34216.293 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Komagataella phaffii (菌類) / 参照: UniProt: C4R7L2 |
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#4: タンパク質 | 分子量: 20622.980 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Komagataella phaffii (菌類) / 参照: UniProt: C4R2U9 |
#7: タンパク質 | 分子量: 18802.625 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Komagataella phaffii (菌類) / 参照: UniProt: C4R9A1 |
#11: タンパク質 | 分子量: 13832.896 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Komagataella phaffii (菌類) / 参照: UniProt: C4R3Z5 |
-DNA-directed RNA polymerases I, II, and III subunit ... , 2種, 2分子 EH
#5: タンパク質 | 分子量: 24962.680 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Komagataella phaffii (菌類) / 参照: UniProt: C4R3P8 |
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#8: タンパク質 | 分子量: 16249.220 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Komagataella phaffii (菌類) / 参照: UniProt: C4R273 |
-タンパク質 , 7種, 11分子 FMOaebfcgdh
#6: タンパク質 | 分子量: 17803.588 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Komagataella phaffii (菌類) / 参照: UniProt: C4R1V1 | ||||||
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#13: タンパク質 | 分子量: 12606.896 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Komagataella phaffii (菌類) / 発現宿主: Escherichia coli (大腸菌) / 参照: UniProt: C4QZ45 | ||||||
#15: タンパク質 | 分子量: 125503.984 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Komagataella phaffii (菌類) 株: ATCC 76273 / CBS 7435 / CECT 11047 / NRRL Y-11430 / Wegner 21-1 遺伝子: RAD26, PP7435_Chr2-0651 / 発現宿主: Escherichia coli (大腸菌) / 参照: UniProt: F2QSG0 | ||||||
#18: タンパク質 | 分子量: 15735.445 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 発現宿主: Escherichia coli (大腸菌) #19: タンパク質 | 分子量: 11676.703 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: H4C1 / 発現宿主: Escherichia coli (大腸菌) / 参照: UniProt: P62805 #20: タンパク質 | 分子量: 14447.825 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: H2AC4, H2AFM, HIST1H2AB, H2AC8, H2AFA, HIST1H2AE / 発現宿主: Escherichia coli (大腸菌) / 参照: UniProt: P04908 #21: タンパク質 | 分子量: 14217.516 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: H2BC11, H2BFR, HIST1H2BJ / 発現宿主: Escherichia coli (大腸菌) / 参照: UniProt: P06899 |
-RNA polymerase subunit ABC10- ... , 2種, 2分子 JL
#10: タンパク質 | 分子量: 8554.064 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Komagataella phaffii (菌類) / 参照: UniProt: C4R009 |
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#12: タンパク質 | 分子量: 7862.048 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Komagataella phaffii (菌類) / 参照: UniProt: F2QMI1 |
-DNA鎖 , 2種, 2分子 NT
#14: DNA鎖 | 分子量: 61381.906 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) synthetic construct (人工物) |
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#17: DNA鎖 | 分子量: 60845.949 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) synthetic construct (人工物) |
-RNA鎖 , 1種, 1分子 P
#16: RNA鎖 | 分子量: 4969.832 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) synthetic construct (人工物) |
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-非ポリマー , 2種, 10分子
#22: 化合物 | ChemComp-ZN / #23: 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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-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 |
-試料調製
構成要素 |
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由来(天然) |
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由来(組換発現) |
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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 |
-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: FEI TITAN KRIOS |
電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: FLOOD BEAM |
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: 57.6 e/Å2 フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) |
-解析
ソフトウェア |
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CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
3次元再構成 | 解像度: 6.95 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 29751 / 対称性のタイプ: POINT | ||||||||||||||||||||||||
精密化 | 交差検証法: NONE 立体化学のターゲット値: GeoStd + Monomer Library + CDL v1.2 | ||||||||||||||||||||||||
原子変位パラメータ | Biso mean: 612.64 Å2 | ||||||||||||||||||||||||
拘束条件 |
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