+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-8480 | |||||||||
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タイトル | Cryo-EM map of the Mediator-RNAPII complex | |||||||||
マップデータ | Mediator-RNAPII complex | |||||||||
試料 |
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機能・相同性 | 機能・相同性情報 co-transcriptional lncRNA 3' end processing, cleavage and polyadenylation pathway / RNA Polymerase I Transcription Initiation / RNA polymerase II transcribes snRNA genes / Processing of Capped Intron-Containing Pre-mRNA / 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 / RNA Polymerase III Transcription Initiation From Type 1 Promoter / RNA Polymerase III Transcription Initiation From Type 2 Promoter ...co-transcriptional lncRNA 3' end processing, cleavage and polyadenylation pathway / RNA Polymerase I Transcription Initiation / RNA polymerase II transcribes snRNA genes / Processing of Capped Intron-Containing Pre-mRNA / 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 / RNA Polymerase III Transcription Initiation From Type 1 Promoter / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA polymerase II, holoenzyme / mRNA Capping / RNA Pol II CTD phosphorylation and interaction with CE / Estrogen-dependent gene expression / Formation of the Early Elongation Complex / RNA Polymerase I Promoter Escape / RNA Polymerase II Pre-transcription Events / TP53 Regulates Transcription of DNA Repair Genes / Transcriptional regulation by small RNAs / Formation of TC-NER Pre-Incision Complex / Dual incision in TC-NER / Gap-filling DNA repair synthesis and ligation in TC-NER / regulation of septum digestion after cytokinesis / siRNA-mediated pericentric heterochromatin formation / DNA-templated transcription elongation / intracellular phosphate ion homeostasis / core mediator complex / chromatin-protein adaptor activity / mediator complex / termination of RNA polymerase II transcription / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / transcription by RNA polymerase I / nuclear-transcribed mRNA catabolic process / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / transcription by RNA polymerase III / RNA polymerase II activity / 7-methylguanosine mRNA capping / transcription elongation by RNA polymerase I / positive regulation of transcription initiation by RNA polymerase II / transcription-coupled nucleotide-excision repair / tRNA transcription by RNA polymerase III / RNA polymerase I complex / RNA polymerase III complex / positive regulation of translational initiation / RNA polymerase II, core complex / pericentric heterochromatin / translation initiation factor binding / transcription initiation at RNA polymerase II promoter / transcription elongation by RNA polymerase II / transcription coregulator activity / P-body / euchromatin / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / single-stranded DNA binding / transcription by RNA polymerase II / nucleic acid binding / transcription coactivator activity / single-stranded RNA binding / protein dimerization activity / RNA polymerase II cis-regulatory region sequence-specific DNA binding / nucleotide binding / chromatin / regulation of transcription by RNA polymerase II / nucleolus / DNA binding / RNA binding / zinc ion binding / nucleus / metal ion binding / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | Schizosaccharomyces pombe (分裂酵母) / Fission yeast (分裂酵母) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 7.8 Å | |||||||||
データ登録者 | Tsai K-L / Yu X / Gopalan S / Chao T-C / Zhang Y / Florens L / Washburn MP / Murakami K / Conaway RC / Conaway JW / Asturias F | |||||||||
資金援助 | 米国, 2件
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引用 | ジャーナル: Nature / 年: 2017 タイトル: Mediator structure and rearrangements required for holoenzyme formation. 著者: Kuang-Lei Tsai / Xiaodi Yu / Sneha Gopalan / Ti-Chun Chao / Ying Zhang / Laurence Florens / Michael P Washburn / Kenji Murakami / Ronald C Conaway / Joan W Conaway / Francisco J Asturias / 要旨: The conserved Mediator co-activator complex has an essential role in the regulation of RNA polymerase II transcription in all eukaryotes. Understanding the structure and interactions of Mediator is ...The conserved Mediator co-activator complex has an essential role in the regulation of RNA polymerase II transcription in all eukaryotes. Understanding the structure and interactions of Mediator is crucial for determining how the complex influences transcription initiation and conveys regulatory information to the basal transcription machinery. Here we present a 4.4 Å resolution cryo-electron microscopy map of Schizosaccharomyces pombe Mediator in which conserved Mediator subunits are individually resolved. The essential Med14 subunit works as a central backbone that connects the Mediator head, middle and tail modules. Comparison with a 7.8 Å resolution cryo-electron microscopy map of a Mediator-RNA polymerase II holoenzyme reveals that changes in the structure of Med14 facilitate a large-scale Mediator rearrangement that is essential for holoenzyme formation. Our study suggests that access to different conformations and crosstalk between structural elements are essential for the Mediator regulation mechanism, and could explain the capacity of the complex to integrate multiple regulatory signals. | |||||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | EMマップ: SurfViewMolmilJmol/JSmol |
添付画像 |
-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_8480.map.gz | 19.4 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-8480-v30.xml emd-8480.xml | 43.3 KB 43.3 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_8480_fsc_1.xml emd_8480_fsc_2.xml | 6.3 KB 6.3 KB | 表示 表示 | FSCデータファイル |
画像 | emd_8480.png | 275.7 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-8480 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-8480 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_8480_validation.pdf.gz | 476.8 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_8480_full_validation.pdf.gz | 476.3 KB | 表示 | |
XML形式データ | emd_8480_validation.xml.gz | 5.6 KB | 表示 | |
CIF形式データ | emd_8480_validation.cif.gz | 6.3 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8480 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8480 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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「今月の分子」の関連する項目 |
-マップ
ファイル | ダウンロード / ファイル: emd_8480.map.gz / 形式: CCP4 / 大きさ: 20.8 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Mediator-RNAPII complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 2.62 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
-試料の構成要素
+全体 : complex of Mediator and RNAPII
+超分子 #1: complex of Mediator and RNAPII
+分子 #1: Mediator complex subunit 6
+分子 #2: Mediator complex subunit 8
+分子 #3: Mediator complex subunit 17
+分子 #4: Mediator complex subunit 18
+分子 #5: Mediator complex subunit 20
+分子 #6: Mediator complex subunit 11
+分子 #7: Mediator complex subunit 22
+分子 #8: Mediator complex subunit 14
+分子 #9: Mediator complex subunit 4
+分子 #10: Mediator complex subunit 7
+分子 #11: Mediator complex subunit 21
+分子 #12: Mediator complex subunit 31
+分子 #13: Mediator complex subunit 27
+分子 #14: Mediator complex subunit 9
+分子 #15: Mediator complex subunit 19
+分子 #16: Mediator complex subunit 10
+分子 #17: RNA polymerase II subunit Rpb1
+分子 #18: RNA polymerase II subunit Rpb2
+分子 #19: RNA polymerase II subunit Rpb3
+分子 #20: RNA polymerase II subunit Rpb4
+分子 #21: RNA polymerase II subunit Rpb5
+分子 #22: RNA polymerase II subunit Rpb6
+分子 #23: RNA polymerase II subunit Rpb7
+分子 #24: RNA polymerase II subunit Rpb8
+分子 #25: RNA polymerase II subunit Rpb9
+分子 #26: RNA polymerase II subunit Rpb10
+分子 #27: RNA polymerase II subunit Rpb11
+分子 #28: RNA polymerase II subunit Rpb12
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
濃度 | 1 mg/mL |
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緩衝液 | pH: 7.4 詳細: 20 mM HEPES, pH 7.4, 200 mM potassium acetate, 5 mM b-Mercaptoethanol, 0.01% NP-40 |
グリッド | モデル: C-flat-2/2 4C / 材質: COPPER / メッシュ: 400 |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / 装置: HOMEMADE PLUNGER |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: COUNTING / 平均露光時間: 7.0 sec. / 平均電子線量: 9.6 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | C2レンズ絞り径: 100.0 µm / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm / 最大 デフォーカス(公称値): 4.0 µm / 最小 デフォーカス(公称値): 1.0 µm / 倍率(公称値): 22500 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |