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基本情報
登録情報 | データベース: EMDB / ID: EMD-3178 | |||||||||
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タイトル | Cryo-EM structure of yeast RNA polymerase III elongation complex at 3.9 A | |||||||||
![]() | Reconstruction of elongating RNA polymerase III | |||||||||
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![]() | RNA polymerase III / pol III / transcription / RNA polymerase | |||||||||
機能・相同性 | ![]() RNA Polymerase I Transcription Initiation / Processing of Capped Intron-Containing Pre-mRNA / 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 / RNA Polymerase II Promoter Escape / RNA Polymerase II Transcription Pre-Initiation And Promoter Opening ...RNA Polymerase I Transcription Initiation / Processing of Capped Intron-Containing Pre-mRNA / 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 / 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-templated transcription / RNA Polymerase II Pre-transcription Events / termination of RNA polymerase III transcription / Formation of TC-NER Pre-Incision Complex / termination of RNA polymerase I transcription / RNA polymerase III activity / transcription initiation at RNA polymerase III promoter / RNA Polymerase I Promoter Escape / 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 / transcription by RNA polymerase I / Estrogen-dependent gene expression / transcription by RNA polymerase III / Dual incision in TC-NER / transcription elongation by RNA polymerase I / tRNA transcription by RNA polymerase III / RNA polymerase I activity / RNA polymerase I complex / RNA polymerase III complex / RNA polymerase II, core complex / nucleotidyltransferase activity / transcription initiation at RNA polymerase II promoter / transcription elongation by RNA polymerase II / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / ribosome biogenesis / peroxisome / single-stranded DNA binding / transcription by RNA polymerase II / nucleic acid binding / protein dimerization activity / nucleotide binding / nucleolus / mitochondrion / DNA binding / zinc ion binding / nucleoplasm / nucleus / metal ion binding / cytosol / cytoplasm 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: 3.9 Å | |||||||||
![]() | Hoffmann NA / Jakobi AJ / Moreno-Morcillo M / Glatt S / Kosinski J / Hagen WJ / Sachse C / Muller CW | |||||||||
![]() | ![]() タイトル: Molecular structures of unbound and transcribing RNA polymerase III. 著者: Niklas A Hoffmann / Arjen J Jakobi / María Moreno-Morcillo / Sebastian Glatt / Jan Kosinski / Wim J H Hagen / Carsten Sachse / Christoph W Müller / ![]() 要旨: Transcription of genes encoding small structured RNAs such as transfer RNAs, spliceosomal U6 small nuclear RNA and ribosomal 5S RNA is carried out by RNA polymerase III (Pol III), the largest yet ...Transcription of genes encoding small structured RNAs such as transfer RNAs, spliceosomal U6 small nuclear RNA and ribosomal 5S RNA is carried out by RNA polymerase III (Pol III), the largest yet structurally least characterized eukaryotic RNA polymerase. Here we present the cryo-electron microscopy structures of the Saccharomyces cerevisiae Pol III elongating complex at 3.9 Å resolution and the apo Pol III enzyme in two different conformations at 4.6 and 4.7 Å resolution, respectively, which allow the building of a 17-subunit atomic model of Pol III. The reconstructions reveal the precise orientation of the C82-C34-C31 heterotrimer in close proximity to the stalk. The C53-C37 heterodimer positions residues involved in transcription termination close to the non-template DNA strand. In the apo Pol III structures, the stalk adopts different orientations coupled with closed and open conformations of the clamp. Our results provide novel insights into Pol III-specific transcription and the adaptation of Pol III towards its small transcriptional targets. | |||||||||
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 | ![]() | 11.3 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 9.5 KB 9.5 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 10.3 KB | 表示 | ![]() |
画像 | ![]() | 79.3 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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注釈 | Reconstruction of elongating RNA polymerase III | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.084 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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試料の構成要素
-全体 : S.cerevisiae RNA polymerase III elongation complex at 3.9 A
全体 | 名称: S.cerevisiae RNA polymerase III elongation complex at 3.9 A |
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要素 |
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-超分子 #1000: S.cerevisiae RNA polymerase III elongation complex at 3.9 A
超分子 | 名称: S.cerevisiae RNA polymerase III elongation complex at 3.9 A タイプ: sample / ID: 1000 / 集合状態: Monomer / Number unique components: 20 |
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分子量 | 理論値: 720 KDa |
-分子 #1: RNA polymerase III
分子 | 名称: RNA polymerase III / タイプ: protein_or_peptide / ID: 1 / コピー数: 1 / 集合状態: Monomer / 組換発現: No |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 実験値: 690 KDa |
-分子 #2: Transcription Bubble
分子 | 名称: Transcription Bubble / タイプ: other / ID: 2 / Name.synonym: template DNA, non-template DNA, RNA / 分類: DNA/RNA / Structure: OTHER / Synthetic?: Yes |
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由来(天然) | 生物種: synthetic construct (人工物) |
分子量 | 実験値: 290 KDa |
-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 | 0.2 mg/mL |
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緩衝液 | pH: 7.5 / 詳細: 150mM (NH4)2SO4, 15mM Tris, 10mM DTT, 5mM MgCl2 |
グリッド | 詳細: Cu 400 mesh Quantifoil 1.2/1.3 |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 95 % / 装置: FEI VITROBOT MARK III 手法: 2.5 ul of sample was applied 15 seconds wait time Blot for 9 seconds before plunging |
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電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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日付 | 2014年10月15日 |
撮影 | カテゴリ: CCD フィルム・検出器のモデル: FEI FALCON II (4k x 4k) 実像数: 2639 / 平均電子線量: 45 e/Å2 詳細: Seven frames were recorded with a dose of 6.5 e/A2 per frame. |
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): 4.2 µm / 最小 デフォーカス(公称値): 1.4 µm / 倍率(公称値): 75000 |
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 |