+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-2231 | |||||||||
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タイトル | The architecture of human general transcription factor TFIID core complex | |||||||||
マップデータ | Cryo-Em structure of the recombinant human 7TAF complex | |||||||||
試料 |
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キーワード | human TFIID / general transcription factor / transcription / multibac / recombinant protein | |||||||||
機能・相同性 | 機能・相同性情報 SAGA complex => GO:0000124 / SAGA complex => GO:0000124 / : / transcription cis-regulatory region binding => GO:0000976 / : / modulation by virus of host process / protein binding / transcription factor TFTC complex / transcription factor TFIID complex / viral process ...SAGA complex => GO:0000124 / SAGA complex => GO:0000124 / : / transcription cis-regulatory region binding => GO:0000976 / : / modulation by virus of host process / protein binding / transcription factor TFTC complex / transcription factor TFIID complex / viral process / transcription initiation at RNA polymerase II promoter / transcription elongation by RNA polymerase II / positive regulation of DNA-binding transcription factor activity / actin cytoskeleton / transcription coactivator activity / protein heterodimerization activity / DNA-binding transcription factor activity / DNA-templated transcription / nucleolus / perinuclear region of cytoplasm / DNA binding / nucleus 類似検索 - 分子機能 | |||||||||
生物種 | Homo sapiens (ヒト) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 14.3 Å | |||||||||
データ登録者 | Bieniossek C / Papai G / Schaffitzel C / Garzoni F / Chaillet M / Scheer E / Papadopoulos P / Tora L / Schultz P / Berger I | |||||||||
引用 | ジャーナル: Nature / 年: 2013 タイトル: The architecture of human general transcription factor TFIID core complex. 著者: Christoph Bieniossek / Gabor Papai / Christiane Schaffitzel / Frederic Garzoni / Maxime Chaillet / Elisabeth Scheer / Petros Papadopoulos / Laszlo Tora / Patrick Schultz / Imre Berger / 要旨: The initiation of gene transcription by RNA polymerase II is regulated by a plethora of proteins in human cells. The first general transcription factor to bind gene promoters is transcription factor ...The initiation of gene transcription by RNA polymerase II is regulated by a plethora of proteins in human cells. The first general transcription factor to bind gene promoters is transcription factor IID (TFIID). TFIID triggers pre-initiation complex formation, functions as a coactivator by interacting with transcriptional activators and reads epigenetic marks. TFIID is a megadalton-sized multiprotein complex composed of TATA-box-binding protein (TBP) and 13 TBP-associated factors (TAFs). Despite its crucial role, the detailed architecture and assembly mechanism of TFIID remain elusive. Histone fold domains are prevalent in TAFs, and histone-like tetramer and octamer structures have been proposed in TFIID. A functional core-TFIID subcomplex was revealed in Drosophila nuclei, consisting of a subset of TAFs (TAF4, TAF5, TAF6, TAF9 and TAF12). These core subunits are thought to be present in two copies in holo-TFIID, in contrast to TBP and other TAFs that are present in a single copy, conveying a transition from symmetry to asymmetry in the TFIID assembly pathway. Here we present the structure of human core-TFIID determined by cryo-electron microscopy at 11.6 Å resolution. Our structure reveals a two-fold symmetric, interlaced architecture, with pronounced protrusions, that accommodates all conserved structural features of the TAFs including the histone folds. We further demonstrate that binding of one TAF8-TAF10 complex breaks the original symmetry of core-TFIID. We propose that the resulting asymmetric structure serves as a functional scaffold to nucleate holo-TFIID assembly, by accreting one copy each of the remaining TAFs and TBP. | |||||||||
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_2231.map.gz | 1.7 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-2231-v30.xml emd-2231.xml | 22.3 KB 22.3 KB | 表示 表示 | EMDBヘッダ |
画像 | EMD-2231-7TAF-front.png | 104.1 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-2231 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-2231 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_2231_validation.pdf.gz | 188.8 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_2231_full_validation.pdf.gz | 187.9 KB | 表示 | |
XML形式データ | emd_2231_validation.xml.gz | 6 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-2231 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-2231 | 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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-マップ
ファイル | ダウンロード / ファイル: emd_2231.map.gz / 形式: CCP4 / 大きさ: 15.3 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Cryo-Em structure of the recombinant human 7TAF 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: 1.87 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 : Recombinant human 7TAF complex containing TAF4, TAF5, TAF6,, TAF8...
全体 | 名称: Recombinant human 7TAF complex containing TAF4, TAF5, TAF6,, TAF8, TAF9, TAF10 and TAF12. |
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要素 |
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-超分子 #1000: Recombinant human 7TAF complex containing TAF4, TAF5, TAF6,, TAF8...
超分子 | 名称: Recombinant human 7TAF complex containing TAF4, TAF5, TAF6,, TAF8, TAF9, TAF10 and TAF12. タイプ: sample / ID: 1000 / 詳細: The sample was fixed by GraFix. / 集合状態: Monomer / Number unique components: 7 |
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分子量 | 理論値: 700 KDa |
-分子 #1: TATA box binding protein (TBP)-associated factor 5
分子 | 名称: TATA box binding protein (TBP)-associated factor 5 / タイプ: protein_or_peptide / ID: 1 / Name.synonym: TAF5 / コピー数: 2 / 集合状態: Dimer / 組換発現: Yes |
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由来(天然) | 生物種: Homo sapiens (ヒト) / 別称: Human / 細胞中の位置: Nucleus |
分子量 | 理論値: 100 KDa |
組換発現 | 生物種: Baculovirus EMBacY / 組換プラスミド: pTF |
配列 | GO: actin cytoskeleton, transcription factor TFIID complex, transcription factor TFTC complex, DNA-binding transcription factor activity, transcription cis-regulatory region binding => GO:0000976, GO: ...GO: actin cytoskeleton, transcription factor TFIID complex, transcription factor TFTC complex, DNA-binding transcription factor activity, transcription cis-regulatory region binding => GO:0000976, GO: 0016573, transcription elongation by RNA polymerase II, transcription initiation at RNA polymerase II promoter, viral process, modulation by virus of host process InterPro: WD40 repeat, LIS1 homology motif, TFIID subunit TAF5, NTD2 domain, WD40/YVTN repeat-like-containing domain superfamily, INTERPRO: IPR017986, WD40 repeat, conserved site, G-protein beta WD-40 repeat |
-分子 #2: TATA box binding protein (TBP)-associated factor 6
分子 | 名称: TATA box binding protein (TBP)-associated factor 6 / タイプ: protein_or_peptide / ID: 2 / Name.synonym: TAF6 / コピー数: 2 / 集合状態: Dimer / 組換発現: Yes |
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由来(天然) | 生物種: Homo sapiens (ヒト) / 別称: Human / 細胞中の位置: Nucleus |
分子量 | 理論値: 70 KDa |
組換発現 | 生物種: Baculovirus EMBacY / 組換プラスミド: pTF |
配列 | GO: actin cytoskeleton, transcription factor TFIID complex, transcription factor TFTC complex, DNA-binding transcription factor activity, transcription cis-regulatory region binding => GO:0000976, GO: ...GO: actin cytoskeleton, transcription factor TFIID complex, transcription factor TFTC complex, DNA-binding transcription factor activity, transcription cis-regulatory region binding => GO:0000976, GO: 0016573, transcription elongation by RNA polymerase II, transcription initiation at RNA polymerase II promoter, viral process, modulation by virus of host process InterPro: TATA box binding protein associated factor (TAF), histone-like fold domain, Histone-fold, TAF6, C-terminal HEAT repeat domain |
-分子 #3: TATA box binding protein (TBP)-associated factor 9
分子 | 名称: TATA box binding protein (TBP)-associated factor 9 / タイプ: protein_or_peptide / ID: 3 / Name.synonym: TAF9 / コピー数: 2 / 集合状態: Dimer / 組換発現: Yes |
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由来(天然) | 生物種: Homo sapiens (ヒト) / 別称: Human / 細胞中の位置: Nucleus |
分子量 | 理論値: 32 KDa |
組換発現 | 生物種: Baculovirus EMBacY / 組換プラスミド: pTF |
配列 | GO: actin cytoskeleton, transcription factor TFIID complex, transcription factor TFTC complex, DNA-binding transcription factor activity, transcription cis-regulatory region binding => GO:0000976, GO: ...GO: actin cytoskeleton, transcription factor TFIID complex, transcription factor TFTC complex, DNA-binding transcription factor activity, transcription cis-regulatory region binding => GO:0000976, GO: 0016573, transcription elongation by RNA polymerase II, transcription initiation at RNA polymerase II promoter, viral process, modulation by virus of host process InterPro: Transcription initiation factor TAFII31, Histone-fold |
-分子 #4: TATA box binding protein (TBP)-associated factor 4
分子 | 名称: TATA box binding protein (TBP)-associated factor 4 / タイプ: protein_or_peptide / ID: 4 / Name.synonym: TAF4 / コピー数: 2 / 集合状態: Dimer / 組換発現: Yes |
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由来(天然) | 生物種: Homo sapiens (ヒト) / 別称: Human / 細胞中の位置: Nucleus |
分子量 | 理論値: 135 KDa |
組換発現 | 生物種: Baculovirus EMBacY / 組換プラスミド: pTF |
配列 | GO: actin cytoskeleton, transcription factor TFIID complex, transcription factor TFTC complex, DNA-binding transcription factor activity, transcription cis-regulatory region binding => GO:0000976, GO: ...GO: actin cytoskeleton, transcription factor TFIID complex, transcription factor TFTC complex, DNA-binding transcription factor activity, transcription cis-regulatory region binding => GO:0000976, GO: 0016573, transcription elongation by RNA polymerase II, transcription initiation at RNA polymerase II promoter, viral process, modulation by virus of host process InterPro: TAFH/NHR1, Transcription initiation factor TFIID component TAF4, C-terminal, Histone-fold |
-分子 #5: TATA box binding protein (TBP)-associated factor 12
分子 | 名称: TATA box binding protein (TBP)-associated factor 12 / タイプ: protein_or_peptide / ID: 5 / Name.synonym: TAF12 / コピー数: 2 / 集合状態: Dimer / 組換発現: Yes |
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由来(天然) | 生物種: Homo sapiens (ヒト) / 別称: Human / 細胞中の位置: Nucleus |
分子量 | 理論値: 15 KDa |
組換発現 | 生物種: Baculovirus EMBacY / 組換プラスミド: pTF |
配列 | GO: SAGA complex => GO:0000124, SAGA complex => GO:0000124, transcription factor TFIID complex, transcription factor TFTC complex, DNA binding, transcription coactivator activity, GO: 0043966, ...GO: SAGA complex => GO:0000124, SAGA complex => GO:0000124, transcription factor TFIID complex, transcription factor TFTC complex, DNA binding, transcription coactivator activity, GO: 0043966, positive regulation of DNA-binding transcription factor activity, transcription elongation by RNA polymerase II, transcription initiation at RNA polymerase II promoter, viral process InterPro: Histone-fold, Transcription initiation factor TFIID subunit 12 domain |
-分子 #6: TATA box binding protein (TBP)-associated factor 8
分子 | 名称: TATA box binding protein (TBP)-associated factor 8 / タイプ: protein_or_peptide / ID: 6 / Name.synonym: TAF8 / コピー数: 1 / 集合状態: Monomer / 組換発現: Yes |
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由来(天然) | 生物種: Homo sapiens (ヒト) / 別称: Human / 細胞中の位置: Nucleus |
分子量 | 理論値: 43 KDa |
組換発現 | 生物種: Baculovirus EMBacY / 組換プラスミド: pTF |
配列 | GO: protein heterodimerization activity, protein binding, transcription factor TFIID complex, perinuclear region of cytoplasm, nucleus, nucleolus, DNA-templated transcription InterPro: Bromodomain associated domain, Histone-fold, Transcription factor TFIID, subunit 8, C-terminal |
-分子 #7: TATA box binding protein (TBP)-associated factor 10
分子 | 名称: TATA box binding protein (TBP)-associated factor 10 / タイプ: protein_or_peptide / ID: 7 / Name.synonym: TAF10 / コピー数: 1 / 集合状態: Monomer / 組換発現: Yes |
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由来(天然) | 生物種: Homo sapiens (ヒト) / 別称: Human / 細胞中の位置: Nucleus |
分子量 | 理論値: 43 KDa |
組換発現 | 生物種: Baculovirus EMBacY / 組換プラスミド: pTF |
配列 | GO: protein heterodimerization activity, protein binding, transcription factor TFIID complex, perinuclear region of cytoplasm, nucleus, nucleolus, DNA-templated transcription InterPro: Transcription initiation factor TFIID, 23-30kDa subunit |
-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 | 0.5 mg/mL |
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緩衝液 | pH: 8 / 詳細: 50mM HEPES pH 8.0, 150mM KCl |
グリッド | 詳細: Quantifoil 300 mesh Cu/Rh holey carbon grid R2/2 |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 95 % / 装置: FEI VITROBOT MARK IV / 手法: 4s, blot force 5 |
-電子顕微鏡法
顕微鏡 | FEI POLARA 300 |
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日付 | 2011年6月18日 |
撮影 | カテゴリ: CCD / フィルム・検出器のモデル: GENERIC CCD / デジタル化 - サンプリング間隔: 14.5 µm / 実像数: 170 / 平均電子線量: 16 e/Å2 / ビット/ピクセル: 16 |
電子線 | 加速電圧: 100 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 倍率(補正後): 78120 / 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): 3.6 µm / 最小 デフォーカス(公称値): 1.5 µm / 倍率(公称値): 78000 |
試料ステージ | 試料ホルダーモデル: OTHER |
実験機器 | モデル: Tecnai Polara / 画像提供: FEI Company |
-画像解析
詳細 | Picking was done with e2boxer from EMAN2, first alignment cycles were done in Imagic and refinement in Spider. |
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CTF補正 | 詳細: Each particle |
最終 再構成 | 想定した対称性 - 点群: C1 (非対称) / アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 14.3 Å / 解像度の算出法: FSC 0.5 CUT-OFF / ソフトウェア - 名称: Imagic, Spider / 使用した粒子像数: 44663 |