+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-12450 | |||||||||||||||
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タイトル | RNA Polymerase II-Spt4/5-nucleosome-FACT structure | |||||||||||||||
マップデータ | Map D | |||||||||||||||
試料 |
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キーワード | Chromatin / nucleosome / elongation / histone chaperone / TRANSCRIPTION | |||||||||||||||
機能・相同性 | 機能・相同性情報 Regulation of TP53 Activity through Phosphorylation / regulation of sister chromatid cohesion / FACT complex / regulation of chromatin organization / DSIF complex / nucleosome organization / regulation of transcription elongation by RNA polymerase II / RPB4-RPB7 complex / replication fork protection complex / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay ...Regulation of TP53 Activity through Phosphorylation / regulation of sister chromatid cohesion / FACT complex / regulation of chromatin organization / DSIF complex / nucleosome organization / regulation of transcription elongation by RNA polymerase II / RPB4-RPB7 complex / replication fork protection 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 / transcription elongation-coupled chromatin remodeling / 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 / 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 II transcription / 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 / RNA polymerase II complex binding / transcription initiation at RNA polymerase III promoter / nucleolar large rRNA transcription by RNA polymerase I / Gap-filling DNA repair synthesis and ligation in TC-NER / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / transcription initiation at RNA polymerase I promoter / transcription by RNA polymerase I / Estrogen-dependent gene expression / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / transcription by RNA polymerase III / chromosome, centromeric region / 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 I complex / RNA polymerase III complex / positive regulation of RNA polymerase II transcription preinitiation complex assembly / transcription-coupled nucleotide-excision repair / RNA polymerase III activity / RNA polymerase II, core complex / tRNA transcription by RNA polymerase III / translesion synthesis / RNA polymerase I activity / RNA polymerase II activity / nucleosome binding / translation initiation factor binding / regulation of DNA-templated transcription elongation / DNA-templated transcription initiation / transcription elongation by RNA polymerase II / transcription initiation at RNA polymerase II promoter / P-body / DNA-templated DNA replication / ribonucleoside binding / mRNA processing / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / cytoplasmic stress granule / structural constituent of chromatin / peroxisome / nucleosome / nucleosome assembly / ribosome biogenesis / chromatin organization / chromosome / single-stranded DNA binding / DNA replication / transcription by RNA polymerase II / nucleic acid binding / single-stranded RNA binding / protein dimerization activity / protein heterodimerization activity / DNA repair / nucleotide binding / mRNA binding / chromatin / nucleolus / 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 | Saccharomyces cerevisiae (パン酵母) / Xenopus laevis (アフリカツメガエル) / synthetic RNA (人工物) / synthetic construct (人工物) / Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (パン酵母) | |||||||||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.2 Å | |||||||||||||||
データ登録者 | Farnung L / Ochmann M | |||||||||||||||
資金援助 | European Union, ドイツ, 4件
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引用 | ジャーナル: Nat Struct Mol Biol / 年: 2021 タイトル: Structural basis of nucleosome transcription mediated by Chd1 and FACT. 著者: Lucas Farnung / Moritz Ochmann / Maik Engeholm / Patrick Cramer / 要旨: Efficient transcription of RNA polymerase II (Pol II) through nucleosomes requires the help of various factors. Here we show biochemically that Pol II transcription through a nucleosome is ...Efficient transcription of RNA polymerase II (Pol II) through nucleosomes requires the help of various factors. Here we show biochemically that Pol II transcription through a nucleosome is facilitated by the chromatin remodeler Chd1 and the histone chaperone FACT when the elongation factors Spt4/5 and TFIIS are present. We report cryo-EM structures of transcribing Saccharomyces cerevisiae Pol II-Spt4/5-nucleosome complexes with bound Chd1 or FACT. In the first structure, Pol II transcription exposes the proximal histone H2A-H2B dimer that is bound by Spt5. Pol II has also released the inhibitory DNA-binding region of Chd1 that is poised to pump DNA toward Pol II. In the second structure, Pol II has generated a partially unraveled nucleosome that binds FACT, which excludes Chd1 and Spt5. These results suggest that Pol II progression through a nucleosome activates Chd1, enables FACT binding and eventually triggers transfer of FACT together with histones to upstream DNA. | |||||||||||||||
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_12450.map.gz | 223.4 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-12450-v30.xml emd-12450.xml | 51.6 KB 51.6 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_12450.png | 103.6 KB | ||
Filedesc metadata | emd-12450.cif.gz | 12.6 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-12450 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-12450 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_12450_validation.pdf.gz | 490.7 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_12450_full_validation.pdf.gz | 490.3 KB | 表示 | |
XML形式データ | emd_12450_validation.xml.gz | 6.6 KB | 表示 | |
CIF形式データ | emd_12450_validation.cif.gz | 7.6 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12450 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12450 | 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_12450.map.gz / 形式: CCP4 / 大きさ: 244.1 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Map D | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.05 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 : Complex of RNA polymerase II-Spt4/5-FACT-nucleosome
+超分子 #1: Complex of RNA polymerase II-Spt4/5-FACT-nucleosome
+超分子 #2: RNA (5'-R(P*UP*CP*UP*UP*UP*UP*AP*UP*UP*UP*UP*UP*UP*CP*UP*G)-3')
+超分子 #3: DNA (138-MER)
+超分子 #4: DNA (148-MER)
+超分子 #5: DNA-directed RNA polymerase II subunit RPB1
+超分子 #6: DNA-directed RNA polymerase II subunit RPB2
+超分子 #7: DNA-directed RNA polymerase II subunit RPB3
+超分子 #8: DNA-directed RNA polymerases I, II, and III subunit RPABC1
+超分子 #9: DNA-directed RNA polymerases I, II, and III subunit RPABC2
+超分子 #10: DNA-directed RNA polymerases I, II, and III subunit RPABC3
+超分子 #11: DNA-directed RNA polymerase II subunit RPB9
+超分子 #12: DNA-directed RNA polymerases I, II, and III subunit RPABC5
+超分子 #13: DNA-directed RNA polymerase II subunit RPB11
+超分子 #14: DNA-directed RNA polymerases I, II, and III subunit RPABC4
+超分子 #15: Chromatin elongation factor SPT4
+超分子 #16: Transcription elongation factor SPT5
+超分子 #17: DNA-directed RNA polymerase II subunit RPB4
+超分子 #18: DNA-directed RNA polymerase II subunit RPB7
+超分子 #19: FACT complex subunit POB3
+超分子 #20: FACT complex subunit SPT16
+超分子 #21: Histone H3.2
+超分子 #22: Histone H4
+超分子 #23: Histone H2A type 1
+超分子 #24: Histone H2B 1.1
+分子 #1: RNA (5'-R(P*UP*CP*UP*UP*UP*UP*AP*UP*UP*UP*UP*UP*UP*CP*UP*G)-3')
+分子 #2: DNA (138-MER)
+分子 #3: DNA (148-MER)
+分子 #4: DNA-directed RNA polymerase II subunit RPB1
+分子 #5: DNA-directed RNA polymerase II subunit RPB2
+分子 #6: DNA-directed RNA polymerase II subunit RPB3
+分子 #7: DNA-directed RNA polymerases I, II, and III subunit RPABC1
+分子 #8: DNA-directed RNA polymerases I, II, and III subunit RPABC2
+分子 #9: DNA-directed RNA polymerases I, II, and III subunit RPABC3
+分子 #10: DNA-directed RNA polymerase II subunit RPB9
+分子 #11: DNA-directed RNA polymerases I, II, and III subunit RPABC5
+分子 #12: DNA-directed RNA polymerase II subunit RPB11
+分子 #13: DNA-directed RNA polymerases I, II, and III subunit RPABC4
+分子 #14: Chromatin elongation factor SPT4
+分子 #15: Transcription elongation factor SPT5
+分子 #16: DNA-directed RNA polymerase II subunit RPB4
+分子 #17: DNA-directed RNA polymerase II subunit RPB7
+分子 #18: FACT complex subunit POB3
+分子 #19: FACT complex subunit SPT16
+分子 #20: Histone H3.2
+分子 #21: Histone H4
+分子 #22: Histone H2A type 1
+分子 #23: Histone H2B 1.1
+分子 #24: ZINC ION
+分子 #25: MAGNESIUM ION
-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
緩衝液 | pH: 7.4 |
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グリッド | モデル: Quantifoil R2/2 / 材質: GOLD / メッシュ: 200 |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277 K / 装置: FEI VITROBOT MARK IV |
-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 K3 BIOQUANTUM (6k x 4k) 平均電子線量: 40.25 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: SPOT SCAN / 撮影モード: BRIGHT FIELD |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
-画像解析
初期モデル | モデルのタイプ: OTHER |
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最終 再構成 | 解像度のタイプ: BY AUTHOR / 解像度: 3.2 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 47138 |
初期 角度割当 | タイプ: MAXIMUM LIKELIHOOD |
最終 角度割当 | タイプ: MAXIMUM LIKELIHOOD |