+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-21514 | |||||||||
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タイトル | map of the mammalian Mediator complex | |||||||||
マップデータ | Mammalian Mediator complex | |||||||||
試料 |
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機能・相同性 | 機能・相同性情報 positive regulation of T cell extravasation / negative regulation of smooth muscle cell differentiation / core mediator complex / nuclear retinoic acid receptor binding / mediator complex / limb development / nuclear thyroid hormone receptor binding / cortical actin cytoskeleton / termination of RNA polymerase II transcription / skeletal muscle cell differentiation ...positive regulation of T cell extravasation / negative regulation of smooth muscle cell differentiation / core mediator complex / nuclear retinoic acid receptor binding / mediator complex / limb development / nuclear thyroid hormone receptor binding / cortical actin cytoskeleton / termination of RNA polymerase II transcription / skeletal muscle cell differentiation / somatic stem cell population maintenance / blastocyst development / positive regulation of transcription initiation by RNA polymerase II / nuclear retinoid X receptor binding / negative regulation of fibroblast proliferation / ubiquitin ligase complex / histone acetyltransferase activity / RNA polymerase II preinitiation complex assembly / nuclear receptor coactivator activity / positive regulation of transcription elongation by RNA polymerase II / transcription coregulator activity / transcription coactivator binding / ubiquitin protein ligase activity / actin binding / transcription regulator complex / transcription by RNA polymerase II / transcription coactivator activity / cytoskeleton / nuclear body / protein ubiquitination / RNA polymerase II cis-regulatory region sequence-specific DNA binding / protein-containing complex binding / positive regulation of gene expression / regulation of DNA-templated transcription / nucleolus / regulation of transcription by RNA polymerase II / positive regulation of DNA-templated transcription / positive regulation of transcription by RNA polymerase II / DNA binding / nucleoplasm / membrane / nucleus / cytoplasm / cytosol 類似検索 - 分子機能 | |||||||||
生物種 | Mus musculus (ハツカネズミ) / Mouse (ネズミ) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 4.02 Å | |||||||||
データ登録者 | Zhao H / Young N / Asturias F | |||||||||
資金援助 | 米国, 2件
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引用 | ジャーナル: Cell / 年: 2019 タイトル: A Pliable Mediator Acts as a Functional Rather Than an Architectural Bridge between Promoters and Enhancers. 著者: Laila El Khattabi / Haiyan Zhao / Jens Kalchschmidt / Natalie Young / Seolkyoung Jung / Peter Van Blerkom / Philippe Kieffer-Kwon / Kyong-Rim Kieffer-Kwon / Solji Park / Xiang Wang / Jordan ...著者: Laila El Khattabi / Haiyan Zhao / Jens Kalchschmidt / Natalie Young / Seolkyoung Jung / Peter Van Blerkom / Philippe Kieffer-Kwon / Kyong-Rim Kieffer-Kwon / Solji Park / Xiang Wang / Jordan Krebs / Subhash Tripathi / Noboru Sakabe / Débora R Sobreira / Su-Chen Huang / Suhas S P Rao / Nathanael Pruett / Daniel Chauss / Erica Sadler / Andrea Lopez / Marcelo A Nóbrega / Erez Lieberman Aiden / Francisco J Asturias / Rafael Casellas / 要旨: While Mediator plays a key role in eukaryotic transcription, little is known about its mechanism of action. This study combines CRISPR-Cas9 genetic screens, degron assays, Hi-C, and cryoelectron ...While Mediator plays a key role in eukaryotic transcription, little is known about its mechanism of action. This study combines CRISPR-Cas9 genetic screens, degron assays, Hi-C, and cryoelectron microscopy (cryo-EM) to dissect the function and structure of mammalian Mediator (mMED). Deletion analyses in B, T, and embryonic stem cells (ESC) identified a core of essential subunits required for Pol II recruitment genome-wide. Conversely, loss of non-essential subunits mostly affects promoters linked to multiple enhancers. Contrary to current models, however, mMED and Pol II are dispensable to physically tether regulatory DNA, a topological activity requiring architectural proteins. Cryo-EM analysis revealed a conserved core, with non-essential subunits increasing structural complexity of the tail module, a primary transcription factor target. Changes in tail structure markedly increase Pol II and kinase module interactions. We propose that Mediator's structural pliability enables it to integrate and transmit regulatory signals and act as a functional, rather than an architectural bridge, between promoters and enhancers. | |||||||||
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_21514.map.gz | 621.2 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-21514-v30.xml emd-21514.xml | 44.9 KB 44.9 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_21514_fsc.xml | 21.2 KB | 表示 | FSCデータファイル |
画像 | emd_21514.png | 133.7 KB | ||
その他 | emd_21514_additional_1.map.gz emd_21514_additional_2.map.gz | 527.6 MB 10.6 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-21514 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-21514 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_21514_validation.pdf.gz | 585.6 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_21514_full_validation.pdf.gz | 585.2 KB | 表示 | |
XML形式データ | emd_21514_validation.xml.gz | 16.9 KB | 表示 | |
CIF形式データ | emd_21514_validation.cif.gz | 23.6 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21514 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21514 | 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_21514.map.gz / 形式: CCP4 / 大きさ: 669.9 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Mammalian Mediator complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.11 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
-追加マップ: Mammalian Mediator complex
ファイル | emd_21514_additional_1.map | ||||||||||||
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注釈 | Mammalian Mediator complex | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-追加マップ: Mammalian Mediator complex
ファイル | emd_21514_additional_2.map | ||||||||||||
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注釈 | Mammalian Mediator complex | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-試料の構成要素
+全体 : Mouse mediator complex
+超分子 #1: Mouse mediator complex
+分子 #1: Mediator of RNA polymerase II transcription subunit 1
+分子 #2: Mediator of RNA polymerase II transcription subunit 4
+分子 #3: Mediator of RNA polymerase II transcription subunit 6
+分子 #4: Mediator of RNA polymerase II transcription subunit 7
+分子 #5: Mediator of RNA polymerase II transcription subunit 8
+分子 #6: Mediator of RNA polymerase II transcription subunit 9
+分子 #7: Mediator of RNA polymerase II transcription subunit 10
+分子 #8: Mediator of RNA polymerase II transcription subunit 11
+分子 #9: Mediator of RNA polymerase II transcription subunit 14
+分子 #10: Mediator of RNA polymerase II transcription subunit 15
+分子 #11: Mediator of RNA polymerase II transcription subunit 16
+分子 #12: Mediator of RNA polymerase II transcription subunit 17
+分子 #13: Mediator of RNA polymerase II transcription subunit 18
+分子 #14: Mediator of RNA polymerase II transcription subunit 19
+分子 #15: Mediator of RNA polymerase II transcription subunit 20
+分子 #16: Mediator of RNA polymerase II transcription subunit 21
+分子 #17: Mediator of RNA polymerase II transcription subunit 22
+分子 #18: Mediator of RNA polymerase II transcription subunit 23
+分子 #19: Mediator of RNA polymerase II transcription subunit 24
+分子 #20: Mediator of RNA polymerase II transcription subunit 25
+分子 #21: Mediator of RNA polymerase II transcription subunit 27
+分子 #22: Mediator of RNA polymerase II transcription subunit 28
+分子 #23: Mediator of RNA polymerase II transcription subunit 29
+分子 #24: Mediator of RNA polymerase II transcription subunit 30
+分子 #25: Mediator of RNA polymerase II transcription subunit 31
-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 | 0.1 mg/mL |
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緩衝液 | pH: 7.9 |
糖包埋 | 材質: ice |
グリッド | モデル: PELCO Ultrathin Carbon with Lacey Carbon / 材質: COPPER / メッシュ: 400 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: CONTINUOUS / 支持フィルム - Film thickness: 30.0 nm / 前処理 - タイプ: PLASMA CLEANING |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277 K / 装置: HOMEMADE PLUNGER |
-電子顕微鏡法
顕微鏡 | FEI TALOS ARCTICA |
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撮影 | フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 平均電子線量: 22.5 e/Å2 |
電子線 | 加速電圧: 200 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | C2レンズ絞り径: 50.0 µm / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm |
実験機器 | モデル: Talos Arctica / 画像提供: FEI Company |