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基本情報
登録情報 | データベース: EMDB / ID: EMD-5407 | |||||||||
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タイトル | Cryo-EM map of a yeast minimal preinitiation complex interacting with the Mediator Head module | |||||||||
![]() | Cryo-EM map of a complex including RNA polymerase II, Mediator Head module, TFIIF, TFIIB, TBP, and promoter DNA. TFIIB, TBP, and DNA are not visible in the map and are presumed to be disordered | |||||||||
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![]() | transcription / RNA polymerase II / Mediator / Head module / preinitiation complex | |||||||||
機能・相同性 | ![]() RNA polymerase II complex recruiting activity / core mediator complex / TFIIH-class transcription factor complex binding / negative regulation of ribosomal protein gene transcription by RNA polymerase II / transcription open complex formation at RNA polymerase II promoter / RPB4-RPB7 complex / mediator complex / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / RNA Polymerase I Transcription Initiation / Processing of Capped Intron-Containing Pre-mRNA ...RNA polymerase II complex recruiting activity / core mediator complex / TFIIH-class transcription factor complex binding / negative regulation of ribosomal protein gene transcription by RNA polymerase II / transcription open complex formation at RNA polymerase II promoter / RPB4-RPB7 complex / mediator complex / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / RNA Polymerase I Transcription Initiation / Processing of Capped Intron-Containing Pre-mRNA / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA Pol II CTD phosphorylation and interaction with CE / Formation of the Early Elongation Complex / mRNA Capping / RNA polymerase II transcribes snRNA genes / TP53 Regulates Transcription of DNA Repair Genes / termination of RNA polymerase II transcription / 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-templated transcription / termination of RNA polymerase III transcription / RNA Polymerase II Pre-transcription Events / : / Formation of TC-NER Pre-Incision Complex / : / termination of RNA polymerase I transcription / RNA Polymerase I Promoter Escape / transcription initiation at RNA polymerase III promoter / nucleolar large rRNA transcription by RNA polymerase I / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / Gap-filling DNA repair synthesis and ligation in TC-NER / transcription initiation at RNA polymerase I promoter / transcription by RNA polymerase I / TFIID-class transcription factor complex binding / Estrogen-dependent gene expression / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / transcription by RNA polymerase III / Dual incision in TC-NER / transcription elongation by RNA polymerase I / positive regulation of transcription initiation by RNA polymerase II / positive regulation of translational initiation / RNA polymerase II core promoter sequence-specific DNA binding / RNA polymerase I complex / RNA polymerase III complex / transcription-coupled nucleotide-excision repair / RNA polymerase III activity / tRNA transcription by RNA polymerase III / RNA polymerase II, core complex / RNA polymerase I activity / translesion synthesis / RNA polymerase II activity / RNA polymerase II preinitiation complex assembly / translation initiation factor binding / TBP-class protein binding / DNA-templated transcription initiation / transcription elongation by RNA polymerase II / transcription initiation at RNA polymerase II promoter / positive regulation of transcription elongation by RNA polymerase II / transcription coregulator activity / P-body / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / cytoplasmic stress granule / transcription corepressor activity / peroxisome / ribosome biogenesis / protein-macromolecule adaptor activity / single-stranded DNA binding / DNA-binding transcription factor 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 / protein domain specific binding / nucleotide binding / mRNA binding / nucleolus / regulation of transcription by RNA polymerase II / negative regulation of transcription by RNA polymerase II / positive regulation of transcription by RNA polymerase II / mitochondrion / DNA binding / zinc ion binding / nucleoplasm / nucleus / metal ion binding / cytoplasm 類似検索 - 分子機能 | |||||||||
生物種 | ![]() ![]() | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 16.0 Å | |||||||||
![]() | Cai G / Chaban Y / Imasaki T / Kovacs JA / Calero G / Penczek PA / Takagi Y / Asturias FJ | |||||||||
![]() | ![]() タイトル: Interaction of the mediator head module with RNA polymerase II. 著者: Gang Cai / Yuriy L Chaban / Tsuyoshi Imasaki / Julio A Kovacs / Guillermo Calero / Pawel A Penczek / Yuichiro Takagi / Francisco J Asturias / ![]() 要旨: Mediator, a large (21 polypeptides, MW ∼1 MDa) complex conserved throughout eukaryotes, plays an essential role in control of gene expression by conveying regulatory signals that influence the ...Mediator, a large (21 polypeptides, MW ∼1 MDa) complex conserved throughout eukaryotes, plays an essential role in control of gene expression by conveying regulatory signals that influence the activity of the preinitiation complex. However, the precise mode of interaction between Mediator and RNA polymerase II (RNAPII), and the mechanism of regulation by Mediator remain elusive. We used cryo-electron microscopy and reconstituted in vitro transcription assays to characterize a transcriptionally-active complex including the Mediator Head module and components of a minimum preinitiation complex (RNAPII, TFIIF, TFIIB, TBP, and promoter DNA). Our results reveal how the Head interacts with RNAPII, affecting its conformation and function. | |||||||||
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構造の表示
ムービー |
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構造ビューア | EMマップ: ![]() ![]() ![]() |
添付画像 |
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マップデータ | ![]() | 1.3 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 15.6 KB 15.6 KB | 表示 表示 | ![]() |
画像 | ![]() | 229.8 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 299.3 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 298.9 KB | 表示 | |
XML形式データ | ![]() | 5 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
EMDBのページ | ![]() ![]() |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Cryo-EM map of a complex including RNA polymerase II, Mediator Head module, TFIIF, TFIIB, TBP, and promoter DNA. TFIIB, TBP, and DNA are not visible in the map and are presumed to be disordered | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 3.83 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
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試料の構成要素
-全体 : Complex of RNA polymerase II, Mediator Head module, TFIIF, TFIIB,...
全体 | 名称: Complex of RNA polymerase II, Mediator Head module, TFIIF, TFIIB, TBP, and promoter DNA |
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要素 |
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-超分子 #1000: Complex of RNA polymerase II, Mediator Head module, TFIIF, TFIIB,...
超分子 | 名称: Complex of RNA polymerase II, Mediator Head module, TFIIF, TFIIB, TBP, and promoter DNA タイプ: sample / ID: 1000 詳細: Although biochemical and functional evidence indicates the presence of all 6 original components (RNA polymerase II, Mediator Head, TFIIF, TFIIB, TBP, DNA), only 3 components (RNA polymerase ...詳細: Although biochemical and functional evidence indicates the presence of all 6 original components (RNA polymerase II, Mediator Head, TFIIF, TFIIB, TBP, DNA), only 3 components (RNA polymerase II, Mediator Head, TFIIF) can be clearly identified in the cryo-EM map. Number unique components: 6 |
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分子量 | 理論値: 900 KDa / 手法: sequence |
-分子 #1: RNA polymerase II
分子 | 名称: RNA polymerase II / タイプ: protein_or_peptide / ID: 1 / Name.synonym: pol II / コピー数: 1 / 組換発現: No / データベース: NCBI |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 540 KDa |
-分子 #2: Head module
分子 | 名称: Head module / タイプ: protein_or_peptide / ID: 2 / Name.synonym: Head / コピー数: 1 / 組換発現: Yes |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 240 KDa |
組換発現 | 生物種: ![]() |
-分子 #3: TFIIF
分子 | 名称: TFIIF / タイプ: protein_or_peptide / ID: 3 / Name.synonym: IIF / コピー数: 1 / 組換発現: No / データベース: NCBI |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 150 KDa |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
濃度 | 0.3 mg/mL |
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緩衝液 | pH: 8 / 詳細: 25 mM KCl, 25 mM Tris-HCl, 10 mM DTT |
グリッド | 詳細: 400 mesh Cu/Rh grids, coated with a perforated carbon film and glow discharged in the presence of amylamine |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 95 % / チャンバー内温度: 77 K / 装置: HOMEMADE PLUNGER / 手法: Blot for ~2 sec before plunging |
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電子顕微鏡法
顕微鏡 | FEI TECNAI F20 |
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温度 | 最低: 77 K / 最高: 120 K / 平均: 115 K |
アライメント法 | Legacy - 非点収差: Objective lens astigmatism was corrected at 125,000X magnification |
日付 | 2008年7月30日 |
撮影 | カテゴリ: CCD フィルム・検出器のモデル: TVIPS TEMCAM-F415 (4k x 4k) 実像数: 500 / 平均電子線量: 10 e/Å2 / ビット/ピクセル: 8 |
電子線 | 加速電圧: 120 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.0 mm / 最大 デフォーカス(公称値): 3.8 µm / 最小 デフォーカス(公称値): 0.8 µm / 倍率(公称値): 50000 |
試料ステージ | 試料ホルダーモデル: GATAN LIQUID NITROGEN |
実験機器 | ![]() モデル: Tecnai F20 / 画像提供: FEI Company |
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画像解析
CTF補正 | 詳細: Each CCD frame |
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最終 再構成 | アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 16.0 Å / 解像度の算出法: FSC 0.5 CUT-OFF / ソフトウェア - 名称: Sparx,Spider / 使用した粒子像数: 51000 |
-原子モデル構築 1
初期モデル | PDB ID: Chain - #0 - Chain ID: A / Chain - #1 - Chain ID: B / Chain - #2 - Chain ID: C / Chain - #3 - Chain ID: D / Chain - #4 - Chain ID: E / Chain - #5 - Chain ID: F / Chain - #6 - Chain ID: G / Chain - #7 - Chain ID: H / Chain - #8 - Chain ID: I / Chain - #9 - Chain ID: J / Chain - #10 - Chain ID: K / Chain - #11 - Chain ID: L |
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ソフトウェア | 名称: Monte Carlo Docking Analysis |
詳細 | Protocol: Rigid body fitting of individual RNA polymerase II structural modules. Fitted individual structural modules previously identified by X-ray crystallographic studies of RNA polymerase II under different crystallization conditions |
精密化 | 空間: REAL / プロトコル: RIGID BODY FIT |
得られたモデル | ![]() PDB-3j1n: ![]() PDB-3j1o: |
-原子モデル構築 2
初期モデル | PDB ID: Chain - #0 - Chain ID: A / Chain - #1 - Chain ID: B / Chain - #2 - Chain ID: C / Chain - #3 - Chain ID: D / Chain - #4 - Chain ID: E / Chain - #5 - Chain ID: F / Chain - #6 - Chain ID: G |
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ソフトウェア | 名称: Monte Carlo Docking Analysis |
詳細 | Protocol: Rigid body fitting of individual Mediator Head structural modules. Fitted individual structural modules previously identified by EM and X-ray crystallographic studies of the Head module |
精密化 | 空間: REAL / プロトコル: RIGID BODY FIT |
得られたモデル | ![]() PDB-3j1n: ![]() PDB-3j1o: |