+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-2786 | |||||||||
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タイトル | Architecture of the RNA polymerase II-Mediator core transcription initiation complex | |||||||||
マップデータ | cITC-cMed | |||||||||
試料 |
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キーワード | transcription / transcription initiation / RNA polymerase II / General Transcription Factors / Mediator | |||||||||
機能・相同性 | 機能・相同性情報 meiotic gene conversion / RNA polymerase II complex recruiting activity / TFIIA-class transcription factor complex binding / transcription factor TFIIIB complex / RNA polymerase III preinitiation complex assembly / RNA polymerase III transcription regulatory region sequence-specific DNA binding / TFIIH-class transcription factor complex binding / core mediator complex / regulation of transcription by RNA polymerase III / RNA polymerase I general transcription initiation factor binding ...meiotic gene conversion / RNA polymerase II complex recruiting activity / TFIIA-class transcription factor complex binding / transcription factor TFIIIB complex / RNA polymerase III preinitiation complex assembly / RNA polymerase III transcription regulatory region sequence-specific DNA binding / TFIIH-class transcription factor complex binding / core mediator complex / regulation of transcription by RNA polymerase III / RNA polymerase I general transcription initiation factor binding / negative regulation of ribosomal protein gene transcription by RNA polymerase II / transcription open complex formation at RNA polymerase II promoter / transcriptional start site selection at RNA polymerase II promoter / RPB4-RPB7 complex / transcription factor TFIIF complex / mediator complex / transcription factor TFIIA complex / RNA polymerase I preinitiation complex assembly / transcription preinitiation complex / DNA binding, bending / RNA Polymerase I Transcription Initiation / Processing of Capped Intron-Containing Pre-mRNA / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / 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 / transcription factor TFIID complex / 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 polymerase II general transcription initiation factor activity / RNA-templated transcription / RNA Polymerase II Pre-transcription Events / termination of RNA polymerase III transcription / Formation of TC-NER Pre-Incision Complex / transcription initiation at RNA polymerase III promoter / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / termination of RNA polymerase I transcription / RNA Polymerase I Promoter Escape / nucleolar large rRNA transcription by RNA polymerase I / RNA polymerase II complex binding / Gap-filling DNA repair synthesis and ligation in TC-NER / transcription by RNA polymerase I / transcription initiation at RNA polymerase I promoter / Estrogen-dependent gene expression / TFIID-class transcription factor complex binding / nuclear-transcribed mRNA catabolic process / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / TFIIB-class transcription factor binding / transcription by RNA polymerase III / RNA polymerase II activity / Dual incision in TC-NER / transcription elongation by RNA polymerase I / positive regulation of transcription initiation by RNA polymerase II / transcription-coupled nucleotide-excision repair / tRNA transcription by RNA polymerase III / RNA polymerase II core promoter sequence-specific DNA binding / RNA polymerase I activity / RNA polymerase I complex / RNA polymerase III complex / positive regulation of translational initiation / translesion synthesis / cellular response to nutrient levels / RNA polymerase II, core complex / RNA polymerase II preinitiation complex assembly / translation initiation factor binding / TBP-class protein binding / DNA-templated transcription initiation / transcription initiation at RNA polymerase II promoter / positive regulation of transcription elongation by RNA polymerase II / 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 / mRNA processing / transcription corepressor activity / disordered domain specific binding / peroxisome / ribosome biogenesis / single-stranded DNA binding / cellular response to heat / protein-macromolecule adaptor activity / DNA-binding transcription factor binding / DNA recombination / RNA polymerase II-specific DNA-binding transcription factor binding / transcription regulator complex / 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 類似検索 - 分子機能 | |||||||||
生物種 | Saccharomyces cerevisiae (パン酵母) / Saccharomyces mikatae (酵母) / synthetic construct (人工物) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 9.7 Å | |||||||||
データ登録者 | Plaschka C / Lariviere L / Wenzeck L / Hemann M / Tegunov D / Petrotchenko EV / Borchers CH / Baumeister W / Herzog F / Villa E / Cramer P | |||||||||
引用 | ジャーナル: Nature / 年: 2015 タイトル: Architecture of the RNA polymerase II-Mediator core initiation complex. 著者: C Plaschka / L Larivière / L Wenzeck / M Seizl / M Hemann / D Tegunov / E V Petrotchenko / C H Borchers / W Baumeister / F Herzog / E Villa / P Cramer / 要旨: The conserved co-activator complex Mediator enables regulated transcription initiation by RNA polymerase (Pol) II. Here we reconstitute an active 15-subunit core Mediator (cMed) comprising all ...The conserved co-activator complex Mediator enables regulated transcription initiation by RNA polymerase (Pol) II. Here we reconstitute an active 15-subunit core Mediator (cMed) comprising all essential Mediator subunits from Saccharomyces cerevisiae. The cryo-electron microscopic structure of cMed bound to a core initiation complex was determined at 9.7 Å resolution. cMed binds Pol II around the Rpb4-Rpb7 stalk near the carboxy-terminal domain (CTD). The Mediator head module binds the Pol II dock and the TFIIB ribbon and stabilizes the initiation complex. The Mediator middle module extends to the Pol II foot with a 'plank' that may influence polymerase conformation. The Mediator subunit Med14 forms a 'beam' between the head and middle modules and connects to the tail module that is predicted to bind transcription activators located on upstream DNA. The Mediator 'arm' and 'hook' domains contribute to a 'cradle' that may position the CTD and TFIIH kinase to stimulate Pol II phosphorylation. | |||||||||
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_2786.map.gz | 77.9 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-2786-v30.xml emd-2786.xml | 40.6 KB 40.6 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_2786.png | 97.6 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-2786 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-2786 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_2786_validation.pdf.gz | 235.5 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_2786_full_validation.pdf.gz | 234.5 KB | 表示 | |
XML形式データ | emd_2786_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-2786 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-2786 | 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_2786.map.gz / 形式: CCP4 / 大きさ: 81.8 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | cITC-cMed | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.35 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 : cITC-cMed
+超分子 #1000: cITC-cMed
+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 RPABC 1
+分子 #6: DNA-DIRECTED RNA POLYMERASES I, II, AND III SUBUNIT RPABC 2
+分子 #7: DNA-DIRECTED RNA POLYMERASE II SUBUNIT RPB7
+分子 #8: DNA-DIRECTED RNA POLYMERASES I, II, AND III SUBUNIT RPABC 3
+分子 #9: DNA-DIRECTED RNA POLYMERASE II SUBUNIT RPB9
+分子 #10: DNA-DIRECTED RNA POLYMERASES I, II, AND III SUBUNIT RPABC 5
+分子 #11: DNA-DIRECTED RNA POLYMERASE II SUBUNIT RPB11
+分子 #12: DNA-DIRECTED RNA POLYMERASES I, II, AND III SUBUNIT RPABC 4
+分子 #13: TRANSCRIPTION INITIATION FACTOR IIB
+分子 #14: TATA-box-binding protein
+分子 #15: Transcription initiation factor IIF subunit alpha
+分子 #16: Transcription initiation factor IIF subunit beta
+分子 #17: Mediator of RNA polymerase II transcription subunit 4
+分子 #18: Mediator of RNA polymerase II transcription subunit 6
+分子 #19: Mediator of RNA polymerase II transcription subunit 7
+分子 #20: Mediator of RNA polymerase II transcription subunit 8
+分子 #21: Mediator of RNA polymerase II transcription subunit 9
+分子 #22: Mediator of RNA polymerase II transcription subunit 10
+分子 #23: Mediator of RNA polymerase II transcription subunit 11
+分子 #24: Mediator of RNA polymerase II transcription subunit 14
+分子 #25: Mediator of RNA polymerase II transcription subunit 17
+分子 #26: Mediator of RNA polymerase II transcription subunit 18
+分子 #27: Mediator of RNA polymerase II transcription subunit 19
+分子 #28: Mediator of RNA polymerase II transcription subunit 20
+分子 #29: Mediator of RNA polymerase II transcription subunit 21
+分子 #30: Mediator of RNA polymerase II transcription subunit 22
+分子 #31: Mediator of RNA polymerase II transcription subunit 31
+分子 #32: template DNA
+分子 #33: nontemplate DNA
+分子 #34: RNA
-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 | 0.15 mg/mL |
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緩衝液 | pH: 7.5 詳細: 25 mM HEPES-KOH pH 7.5, 180 mM Potassium acetate, 5 % Glycerol, 5 mM DTT |
凍結 | 凍結剤: ETHANE / 装置: OTHER 手法: Grids were glow-discharged for 20 s before deposition of 4 microliters sample and incubated for 30 s. Grids were washed twice with 4 microliters distilled water, blotted, and then vitrified. |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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撮影 | カテゴリ: CCD / フィルム・検出器のモデル: GATAN K2 (4k x 4k) / 実像数: 2972 / 平均電子線量: 25 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 倍率(補正後): 37169 / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2 mm / 最大 デフォーカス(公称値): 2.5 µm / 最小 デフォーカス(公称値): 1.0 µm / 倍率(公称値): 37000 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
-画像解析
詳細 | Particles were picked using EMAN2 and were processed using RELION 1.2. |
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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: 9.7 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: RELION, 1.2 詳細: The density was filtered according to local resolution and further a temperature factor of minus 340 A2 was applied. 使用した粒子像数: 3267 |