+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-11673 | |||||||||
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タイトル | Cryo-EM structure of human apo RNA Polymerase III | |||||||||
マップデータ | Cryo-EM map of apo human RNA polymerase III, Map A, unsharpened | |||||||||
試料 |
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キーワード | Human RNA polymerase III / Synthesis of short RNAs / apo complex / TRANSCRIPTION | |||||||||
機能・相同性 | 機能・相同性情報 snRNA transcription by RNA polymerase III / RNA Polymerase III Chain Elongation / RNA Polymerase III Transcription Termination / calcitonin gene-related peptide receptor activity / DNA/RNA hybrid binding / regulation of transcription by RNA polymerase I / regulation of transcription by RNA polymerase III / RPAP3/R2TP/prefoldin-like complex / DNA polymerase III complex / Cytosolic sensors of pathogen-associated DNA ...snRNA transcription by RNA polymerase III / RNA Polymerase III Chain Elongation / RNA Polymerase III Transcription Termination / calcitonin gene-related peptide receptor activity / DNA/RNA hybrid binding / regulation of transcription by RNA polymerase I / regulation of transcription by RNA polymerase III / RPAP3/R2TP/prefoldin-like complex / DNA polymerase III complex / Cytosolic sensors of pathogen-associated DNA / RNA Polymerase III Transcription Initiation From Type 1 Promoter / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA Polymerase III Transcription Initiation From Type 3 Promoter / RNA Polymerase III Abortive And Retractive Initiation / positive regulation of innate immune response / Abortive elongation of HIV-1 transcript in the absence of Tat / nucleobase-containing compound metabolic process / RNA Polymerase I Transcription Termination / FGFR2 alternative splicing / MicroRNA (miRNA) biogenesis / Signaling by FGFR2 IIIa TM / Viral Messenger RNA Synthesis / RNA Pol II CTD phosphorylation and interaction with CE during HIV infection / RNA Pol II CTD phosphorylation and interaction with CE / Formation of the Early Elongation Complex / Formation of the HIV-1 Early Elongation Complex / mRNA Capping / PIWI-interacting RNA (piRNA) biogenesis / termination of RNA polymerase III transcription / HIV Transcription Initiation / RNA Polymerase II HIV Promoter Escape / Transcription of the HIV genome / 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 III activity / transcription initiation at RNA polymerase III promoter / mRNA Splicing - Minor Pathway / RNA Polymerase I Transcription Initiation / Pausing and recovery of Tat-mediated HIV elongation / Tat-mediated HIV elongation arrest and recovery / transcription by RNA polymerase I / HIV elongation arrest and recovery / Pausing and recovery of HIV elongation / Processing of Capped Intron-Containing Pre-mRNA / transcription by RNA polymerase III / RNA polymerase II transcribes snRNA genes / neuropeptide signaling pathway / Tat-mediated elongation of the HIV-1 transcript / transcription elongation by RNA polymerase I / Formation of HIV-1 elongation complex containing HIV-1 Tat / tRNA transcription by RNA polymerase III / RNA polymerase I activity / RNA polymerase I complex / RNA polymerase III complex / Formation of HIV elongation complex in the absence of HIV Tat / RNA polymerase II, core complex / RNA Polymerase II Transcription Elongation / Formation of RNA Pol II elongation complex / RNA Polymerase II Pre-transcription Events / Inhibition of DNA recombination at telomere / positive regulation of interferon-beta production / mRNA Splicing - Major Pathway / acrosomal vesicle / protein-DNA complex / TP53 Regulates Transcription of DNA Repair Genes / RNA Polymerase I Promoter Escape / Transcriptional regulation by small RNAs / NoRC negatively regulates rRNA expression / B-WICH complex positively regulates rRNA expression / Transcription-Coupled Nucleotide Excision Repair (TC-NER) / Formation of TC-NER Pre-Incision Complex / ribonucleoside binding / fibrillar center / DNA-directed 5'-3' RNA polymerase activity / Activation of anterior HOX genes in hindbrain development during early embryogenesis / DNA-directed RNA polymerase / Dual incision in TC-NER / Gap-filling DNA repair synthesis and ligation in TC-NER / single-stranded DNA binding / 4 iron, 4 sulfur cluster binding / double-stranded DNA binding / defense response to virus / Estrogen-dependent gene expression / transcription by RNA polymerase II / cell population proliferation / nucleic acid binding / nuclear body / protein dimerization activity / protein stabilization / intracellular membrane-bounded organelle / innate immune response / nucleotide binding / centrosome / DNA-templated transcription / chromatin binding / magnesium ion binding / DNA binding / zinc ion binding 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: 3.3 Å | |||||||||
データ登録者 | Girbig M / Misiaszek AD | |||||||||
引用 | ジャーナル: Nat Struct Mol Biol / 年: 2021 タイトル: Cryo-EM structures of human RNA polymerase III in its unbound and transcribing states. 著者: Mathias Girbig / Agata D Misiaszek / Matthias K Vorländer / Aleix Lafita / Helga Grötsch / Florence Baudin / Alex Bateman / Christoph W Müller / 要旨: RNA polymerase III (Pol III) synthesizes transfer RNAs and other short, essential RNAs. Human Pol III misregulation is linked to tumor transformation, neurodegenerative and developmental disorders, ...RNA polymerase III (Pol III) synthesizes transfer RNAs and other short, essential RNAs. Human Pol III misregulation is linked to tumor transformation, neurodegenerative and developmental disorders, and increased sensitivity to viral infections. Here, we present cryo-electron microscopy structures at 2.8 to 3.3 Å resolution of transcribing and unbound human Pol III. We observe insertion of the TFIIS-like subunit RPC10 into the polymerase funnel, providing insights into how RPC10 triggers transcription termination. Our structures resolve elements absent from Saccharomyces cerevisiae Pol III such as the winged-helix domains of RPC5 and an iron-sulfur cluster, which tethers the heterotrimer subcomplex to the core. The cancer-associated RPC7α isoform binds the polymerase clamp, potentially interfering with Pol III inhibition by tumor suppressor MAF1, which may explain why overexpressed RPC7α enhances tumor transformation. Finally, the human Pol III structure allows mapping of disease-related mutations and may contribute to the development of inhibitors that selectively target Pol III for therapeutic interventions. | |||||||||
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_11673.map.gz | 257.9 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-11673-v30.xml emd-11673.xml | 48.5 KB 48.5 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_11673_fsc.xml | 15.7 KB | 表示 | FSCデータファイル |
画像 | emd_11673.png | 63.8 KB | ||
マスクデータ | emd_11673_msk_1.map | 325 MB | マスクマップ | |
Filedesc metadata | emd-11673.cif.gz | 10.6 KB | ||
その他 | emd_11673_additional_1.map.gz emd_11673_additional_2.map.gz emd_11673_additional_3.map.gz emd_11673_additional_4.map.gz emd_11673_additional_5.map.gz emd_11673_half_map_1.map.gz emd_11673_half_map_2.map.gz | 257.9 MB 241.7 MB 221.8 MB 21.2 MB 9.1 MB 258.8 MB 258.8 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-11673 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-11673 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_11673_validation.pdf.gz | 1 MB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_11673_full_validation.pdf.gz | 1 MB | 表示 | |
XML形式データ | emd_11673_validation.xml.gz | 22.9 KB | 表示 | |
CIF形式データ | emd_11673_validation.cif.gz | 30.4 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-11673 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-11673 | HTTPS FTP |
-関連構造データ
関連構造データ | 7a6hMC 7ae1C 7ae3C 7aeaC C: 同じ文献を引用 (文献) M: このマップから作成された原子モデル |
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類似構造データ | |
電子顕微鏡画像生データ | EMPIAR-10697 (タイトル: Cryo-EM structures of human RNA Polymerase III Data size: 3.3 TB Data #1: LZW-TIFF compressed multiframe micrographs of human RNA Pol III EC [micrographs - multiframe] Data #2: Initially picked particles of human RNA Pol III EC [picked particles - single frame - processed] Data #3: Polished particles of human RNA Pol III EC (final reconstruction) [picked particles - single frame - processed] Data #4: LZW-TIFF compressed multiframe micrographs of human RNA Pol III apo [micrographs - multiframe] Data #5: Initially picked particles of human RNA Pol III apo [picked particles - single frame - processed] Data #6: Polished particles of human RNA Pol III apo (final reconstruction) [picked particles - single frame - processed]) |
-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_11673.map.gz / 形式: CCP4 / 大きさ: 325 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Cryo-EM map of apo human RNA polymerase III, Map A, unsharpened | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 0.822 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
-マスク #1
ファイル | emd_11673_msk_1.map | ||||||||||||
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投影像・断面図 |
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密度ヒストグラム |
-追加マップ: Cryo-EM map of apo human RNA polymerase III,...
ファイル | emd_11673_additional_1.map | ||||||||||||
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注釈 | Cryo-EM map of apo human RNA polymerase III, Map A, locally sharpened with LocalDeblur | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-追加マップ: Cryo-EM map of apo human RNA polymerase III,...
ファイル | emd_11673_additional_2.map | ||||||||||||
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注釈 | Cryo-EM map of apo human RNA polymerase III, Map A1, body 001 from RELION multi-body refinement | ||||||||||||
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密度ヒストグラム |
-追加マップ: Cryo-EM map of apo human RNA polymerase III,...
ファイル | emd_11673_additional_3.map | ||||||||||||
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注釈 | Cryo-EM map of apo human RNA polymerase III, Map A2, body 002 from RELION multi-body refinement, | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-追加マップ: Cryo-EM map of apo human RNA polymerase III,...
ファイル | emd_11673_additional_4.map | ||||||||||||
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注釈 | Cryo-EM map of apo human RNA polymerase III, Map A1, body 001 from RELION multi-body refinement, sharpened via density modification | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-追加マップ: Cryo-EM map of apo human RNA polymerase III,...
ファイル | emd_11673_additional_5.map | ||||||||||||
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注釈 | Cryo-EM map of apo human RNA polymerase III, Map A2, body 002 from RELION multi-body refinement, sharpened via density modification | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: #1
ファイル | emd_11673_half_map_1.map | ||||||||||||
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投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: #2
ファイル | emd_11673_half_map_2.map | ||||||||||||
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投影像・断面図 |
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密度ヒストグラム |
-試料の構成要素
+全体 : Cryo-EM structure of human apo RNA Polymerase III
+超分子 #1: Cryo-EM structure of human apo RNA Polymerase III
+分子 #1: DNA-directed RNA polymerase III subunit RPC1
+分子 #2: DNA-directed RNA polymerase III subunit RPC2
+分子 #3: DNA-directed RNA polymerases I and III subunit RPAC1
+分子 #4: DNA-directed RNA polymerase III subunit RPC9
+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 III subunit RPC8
+分子 #8: DNA-directed RNA polymerases I, II, and III subunit RPABC3
+分子 #9: DNA-directed RNA polymerase III subunit RPC10
+分子 #10: DNA-directed RNA polymerases I, II, and III subunit RPABC5
+分子 #11: DNA-directed RNA polymerases I and III subunit RPAC2
+分子 #12: DNA-directed RNA polymerases I, II, and III subunit RPABC4
+分子 #13: DNA-directed RNA polymerase III subunit RPC5
+分子 #14: DNA-directed RNA polymerase III subunit RPC4
+分子 #15: DNA-directed RNA polymerase III subunit RPC3
+分子 #16: DNA-directed RNA polymerase III subunit RPC6
+分子 #17: DNA-directed RNA polymerase III subunit RPC7
+分子 #18: ZINC ION
+分子 #19: MAGNESIUM ION
+分子 #20: 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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解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
濃度 | 2 mg/mL | ||||||||||||
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緩衝液 | pH: 7.5 構成要素:
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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 / 詳細: Wait time 10 s Blot force 4 Blot time 4 s. |
-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 (6k x 4k) / 検出モード: COUNTING / 平均電子線量: 38.11 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | C2レンズ絞り径: 50.0 µm / 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): 2.25 µm / 最小 デフォーカス(公称値): 0.75 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 |