+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-2229 | |||||||||
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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 3TAF 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 transcription cis-regulatory region binding => GO:0000976 / : / modulation by virus of host process / transcription factor TFTC complex / transcription factor TFIID complex / viral process / transcription initiation at RNA polymerase II promoter / transcription elongation by RNA polymerase II / actin cytoskeleton / DNA-binding transcription factor activity 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: 12.8 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_2229.map.gz | 1.2 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-2229-v30.xml emd-2229.xml | 15 KB 15 KB | 表示 表示 | EMDBヘッダ |
画像 | EMD-2229-3TAF-front.png | 111.8 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-2229 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-2229 | 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_2229.map.gz / 形式: CCP4 / 大きさ: 7.8 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 3TAF complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 2.032 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 3TAF complex containing TAF5, TAF6 and TAF9.
全体 | 名称: Recombinant human 3TAF complex containing TAF5, TAF6 and TAF9. |
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要素 |
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-超分子 #1000: Recombinant human 3TAF complex containing TAF5, TAF6 and TAF9.
超分子 | 名称: Recombinant human 3TAF complex containing TAF5, TAF6 and TAF9. タイプ: sample / ID: 1000 / 詳細: The sample was fixed by GraFix. / 集合状態: Dimer / Number unique components: 3 |
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分子量 | 理論値: 400 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 |
-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 TECNAI 20 |
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日付 | 2010年9月15日 |
撮影 | カテゴリ: FILM / フィルム・検出器のモデル: KODAK SO-163 FILM / デジタル化 - スキャナー: OTHER / デジタル化 - サンプリング間隔: 10.16 µm / 実像数: 16 / 平均電子線量: 16 e/Å2 / ビット/ピクセル: 16 |
電子線 | 加速電圧: 200 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 倍率(補正後): 50012 / 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): 3.45 µm / 最小 デフォーカス(公称値): 1.2 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 |
-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: C2 (2回回転対称) / アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 12.8 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: 11987 |