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基本情報
登録情報 | データベース: EMDB / ID: EMD-3850 | |||||||||||||||
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タイトル | STRUCTURE OF YEAST TRANSCRIPTION PRE-INITIATION COMPLEX WITH TFIIH AND CORE MEDIATOR | |||||||||||||||
![]() | Transcription initiation complex including Pol II, TBP, the transcription factors TFIIA, -B, -E, -F and -H and core Mediator | |||||||||||||||
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![]() | transcription initiation / TRANSCRIPTION / macromolecular complex | |||||||||||||||
機能・相同性 | ![]() meiotic gene conversion / regulation of mitotic recombination / RNA polymerase II promoter clearance / RNA polymerase II complex recruiting activity / transcription open complex formation at RNA polymerase II promoter / TFIIA-class transcription factor complex binding / RNA polymerase III transcription regulatory region sequence-specific DNA binding / TFIIH-class transcription factor complex binding / phosphatidylinositol-5-phosphate binding / RNA polymerase III preinitiation complex assembly ...meiotic gene conversion / regulation of mitotic recombination / RNA polymerase II promoter clearance / RNA polymerase II complex recruiting activity / transcription open complex formation at RNA polymerase II promoter / TFIIA-class transcription factor complex binding / RNA polymerase III transcription regulatory region sequence-specific DNA binding / TFIIH-class transcription factor complex binding / phosphatidylinositol-5-phosphate binding / RNA polymerase III preinitiation complex assembly / transcription factor TFIIIB complex / positive regulation of mitotic recombination / nucleotide-excision repair factor 3 complex / RNA polymerase I general transcription initiation factor binding / core mediator complex / transcription factor TFIIE complex / nucleotide-excision repair, preincision complex assembly / DNA translocase activity / negative regulation of ribosomal protein gene transcription by RNA polymerase II / regulation of transcription by RNA polymerase III / transcription factor TFIIK complex / TFIIF-class transcription factor complex binding / transcriptional start site selection at RNA polymerase II promoter / RPB4-RPB7 complex / transcription factor TFIIF complex / DNA 5'-3' helicase / mediator complex / phosphatidylinositol-3-phosphate binding / transcription factor TFIIA complex / RNA polymerase I preinitiation complex assembly / transcription factor TFIIH core complex / transcription factor TFIIH holo complex / cyclin-dependent protein serine/threonine kinase activator activity / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / DNA binding, bending / RNA Polymerase I Transcription Initiation / transcription preinitiation complex / 3'-5' DNA helicase activity / Processing of Capped Intron-Containing Pre-mRNA / DNA 3'-5' helicase / RNA Polymerase III Transcription Initiation From Type 2 Promoter / poly(A)+ mRNA export from nucleus / RNA Pol II CTD phosphorylation and interaction with CE / Formation of the Early Elongation Complex / mRNA Capping / RNA polymerase II transcribes snRNA genes / termination of RNA polymerase II transcription / TP53 Regulates Transcription of DNA Repair Genes / 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 / termination of RNA polymerase III transcription / RNA polymerase II general transcription initiation factor activity / transcription factor TFIID complex / RNA Polymerase II Pre-transcription Events / RNA-templated transcription / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / Formation of TC-NER Pre-Incision Complex / RNA Polymerase I Promoter Escape / transcription initiation at RNA polymerase III promoter / termination of RNA polymerase I transcription / nucleolar large rRNA transcription by RNA polymerase I / Gap-filling DNA repair synthesis and ligation in TC-NER / transcription initiation at RNA polymerase I promoter / ATPase activator activity / RNA polymerase II complex binding / Estrogen-dependent gene expression / TFIID-class transcription factor complex binding / TFIIB-class transcription factor binding / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / nuclear-transcribed mRNA catabolic process / positive regulation of translational initiation / protein phosphatase activator activity / Dual incision in TC-NER / ATP-dependent activity, acting on DNA / positive regulation of transcription initiation by RNA polymerase II / cellular response to nutrient levels / transcription by RNA polymerase III / translesion synthesis / transcription by RNA polymerase I / RNA polymerase I complex / transcription elongation by RNA polymerase I / RNA polymerase III complex / transcription-coupled nucleotide-excision repair / RNA polymerase II core promoter sequence-specific DNA binding / RNA polymerase II, core complex / DNA helicase activity / tRNA transcription by RNA polymerase III / : / RNA polymerase II preinitiation complex assembly / translation initiation factor binding / TBP-class protein binding / DNA-directed RNA polymerase activity / isomerase activity / nucleotide-excision repair / positive regulation of transcription elongation by RNA polymerase II / transcription coregulator activity / transcription initiation at RNA polymerase II promoter / DNA-templated transcription initiation 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: 5.8 Å | |||||||||||||||
![]() | Schilbach S / Hantsche M / Tegunov D / Dienemann C / Wigge C / Urlaub H / Cramer P | |||||||||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Structures of transcription pre-initiation complex with TFIIH and Mediator. 著者: S Schilbach / M Hantsche / D Tegunov / C Dienemann / C Wigge / H Urlaub / P Cramer / ![]() 要旨: For the initiation of transcription, RNA polymerase II (Pol II) assembles with general transcription factors on promoter DNA to form the pre-initiation complex (PIC). Here we report cryo-electron ...For the initiation of transcription, RNA polymerase II (Pol II) assembles with general transcription factors on promoter DNA to form the pre-initiation complex (PIC). Here we report cryo-electron microscopy structures of the Saccharomyces cerevisiae PIC and PIC-core Mediator complex at nominal resolutions of 4.7 Å and 5.8 Å, respectively. The structures reveal transcription factor IIH (TFIIH), and suggest how the core and kinase TFIIH modules function in the opening of promoter DNA and the phosphorylation of Pol II, respectively. The TFIIH core subunit Ssl2 (a homologue of human XPB) is positioned on downstream DNA by the 'E-bridge' helix in TFIIE, consistent with TFIIE-stimulated DNA opening. The TFIIH kinase module subunit Tfb3 (MAT1 in human) anchors the kinase Kin28 (CDK7), which is mobile in the PIC but preferentially located between the Mediator hook and shoulder in the PIC-core Mediator complex. Open spaces between the Mediator head and middle modules may allow access of the kinase to its substrate, the C-terminal domain of Pol II. | |||||||||||||||
履歴 |
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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: ![]() ![]() ![]() |
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マップデータ | ![]() | 149.6 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 89.6 KB 89.6 KB | 表示 表示 | ![]() |
画像 | ![]() | 193.9 KB | ||
Filedesc metadata | ![]() | 19.6 KB | ||
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「今月の分子」の関連する項目 |
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ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Transcription initiation complex including Pol II, TBP, the transcription factors TFIIA, -B, -E, -F and -H and core Mediator | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.37 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 : Pre-Initiation Complex with core Mediator (PIC-cMed)
+超分子 #1: Pre-Initiation Complex with core Mediator (PIC-cMed)
+超分子 #2: DNA-directed RNA polymerase
+超分子 #3: RNA polymerase II transcription factor B
+超分子 #4: Mediator of RNA polymerase II transcription
+超分子 #5: DNA
+分子 #1: DNA-directed RNA polymerase II subunit RPB1
+分子 #2: DNA-directed RNA polymerase II subunit RPB2
+分子 #3: DNA-directed RNA polymerase II subunit RPB3
+分子 #4: DNA-directed RNA polymerase II subunit RPB4
+分子 #5: DNA-directed RNA polymerases I, II, and III subunit RPABC1
+分子 #6: DNA-directed RNA polymerases I, II, and III subunit RPABC2
+分子 #7: DNA-directed RNA polymerase II subunit RPB7
+分子 #8: DNA-directed RNA polymerases I, II, and III subunit RPABC3
+分子 #9: DNA-directed RNA polymerase II subunit RPB9
+分子 #10: DNA-directed RNA polymerases I, II, and III subunit RPABC5
+分子 #11: DNA-directed RNA polymerase II subunit RPB11
+分子 #12: DNA-directed RNA polymerases I, II, and III subunit RPABC4
+分子 #13: Transcription initiation factor IIB
+分子 #15: TATA-box-binding protein
+分子 #16: Transcription initiation factor IIF subunit alpha
+分子 #17: Transcription initiation factor IIF subunit beta
+分子 #19: Transcription initiation factor IIA large subunit
+分子 #20: Transcription initiation factor IIA subunit 2
+分子 #21: Transcription factor TFIIE subunit
+分子 #22: Transcription initiation factor IIE subunit beta
+分子 #23: General transcription and DNA repair factor IIH helicase subunit XPD
+分子 #24: General transcription and DNA repair factor IIH subunit TFB1
+分子 #25: General transcription and DNA repair factor IIH subunit TFB2
+分子 #26: RNA polymerase II transcription factor B subunit 3
+分子 #27: General transcription and DNA repair factor IIH subunit TFB4
+分子 #28: General transcription and DNA repair factor IIH subunit TFB5
+分子 #29: General transcription and DNA repair factor IIH subunit SSL1
+分子 #30: General transcription and DNA repair factor IIH helicase subunit XPB
+分子 #31: Unknown protein
+分子 #32: Mediator of RNA polymerase II transcription subunit 6
+分子 #33: Mediator of RNA polymerase II transcription subunit 8
+分子 #34: Mediator of RNA polymerase II transcription subunit 11
+分子 #35: Mediator of RNA polymerase II transcription subunit 17
+分子 #36: Mediator of RNA polymerase II transcription subunit 18
+分子 #37: Mediator of RNA polymerase II transcription subunit 20
+分子 #38: Mediator of RNA polymerase II transcription subunit 22
+分子 #39: Mediator of RNA polymerase II transcription subunit 4
+分子 #40: Mediator of RNA polymerase II transcription subunit 7
+分子 #41: Mediator of RNA polymerase II transcription subunit 9
+分子 #42: Mediator of RNA polymerase II transcription subunit 10
+分子 #43: Mediator of RNA polymerase II transcription subunit 14
+分子 #44: Mediator of RNA polymerase II transcription subunit 19
+分子 #45: Mediator of RNA polymerase II transcription subunit 21
+分子 #46: Mediator of RNA polymerase II transcription subunit 31,Mediator o...
+分子 #14: NONTEMPLATE DNA
+分子 #18: TEMPLATE DNA
+分子 #47: ZINC ION
+分子 #48: MAGNESIUM ION
+分子 #49: IRON/SULFUR CLUSTER
-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 | 0.70 mg/mL |
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緩衝液 | pH: 7.6 |
グリッド | モデル: Quantifoil R1.2/1.3 / 材質: GOLD / 前処理 - タイプ: GLOW DISCHARGE 詳細: QUANTIFOIL HOLEY GOLD GRIDS WERE GLOW-DISCHARGED BEFORE DEPOSITION OF SAMPLE. |
凍結 | 凍結剤: ETHANE / 装置: FEI VITROBOT MARK IV 詳細: Grids were blotted for 8 s and plunge-frozen in liquid ethane.. |
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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 / 撮影したグリッド数: 3 / 実像数: 13000 / 平均露光時間: 10.0 sec. / 平均電子線量: 42.0 e/Å2 |
電子線 | 加速電圧: 300 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: 2.7 mm / 最大 デフォーカス(公称値): 5.0 µm / 最小 デフォーカス(公称値): 0.5 µm / 倍率(公称値): 105000 |
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 |
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画像解析
初期モデル | モデルのタイプ: EMDB MAP |
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最終 再構成 | 想定した対称性 - 点群: C1 (非対称) / 解像度のタイプ: BY AUTHOR / 解像度: 5.8 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 16000 |
初期 角度割当 | タイプ: PROJECTION MATCHING / ソフトウェア - 名称: RELION (ver. 2.0.4) |
最終 角度割当 | タイプ: PROJECTION MATCHING / ソフトウェア - 名称: RELION (ver. 2.0.4) |
-原子モデル構築 1
精密化 | 空間: REAL / プロトコル: OTHER |
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得られたモデル | ![]() PDB-5oqm: |