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データを開く
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基本情報
登録情報 | ![]() | |||||||||
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タイトル | Cryo-EM structure of human CST bound to DNA polymerase alpha-primase in a recruitment state | |||||||||
![]() | Human CST-Pol alpha/Primase bound to ssDNA in a recruitment state. | |||||||||
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![]() | Fill-in / Telomere / Replication / DNA BINDING PROTEIN / DNA BINDING PROTEIN-DNA complex | |||||||||
機能・相同性 | ![]() CST complex / positive regulation of DNA primase activity / DNA primase AEP / ribonucleotide binding / telomerase inhibitor activity / DNA replication initiation / DNA/RNA hybrid binding / telomere maintenance via telomere lengthening / Telomere C-strand synthesis initiation / Inhibition of replication initiation of damaged DNA by RB1/E2F1 ...CST complex / positive regulation of DNA primase activity / DNA primase AEP / ribonucleotide binding / telomerase inhibitor activity / DNA replication initiation / DNA/RNA hybrid binding / telomere maintenance via telomere lengthening / Telomere C-strand synthesis initiation / Inhibition of replication initiation of damaged DNA by RB1/E2F1 / alpha DNA polymerase:primase complex / Polymerase switching / regulation of type I interferon production / Processive synthesis on the lagging strand / DNA primase activity / Removal of the Flap Intermediate / single-stranded telomeric DNA binding / Polymerase switching on the C-strand of the telomere / lagging strand elongation / G-rich strand telomeric DNA binding / DNA replication, synthesis of primer / telomere capping / mitotic DNA replication initiation / bone marrow development / intermediate filament cytoskeleton / hematopoietic stem cell proliferation / DNA strand elongation involved in DNA replication / DNA synthesis involved in DNA repair / G1/S-Specific Transcription / telomeric DNA binding / leading strand elongation / DNA replication origin binding / negative regulation of telomere maintenance via telomerase / replicative senescence / Activation of the pre-replicative complex / DNA replication initiation / spleen development / regulation of G2/M transition of mitotic cell cycle / telomere maintenance / thymus development / positive regulation of DNA replication / Defective pyroptosis / multicellular organism growth / fibrillar center / nuclear matrix / double-strand break repair via nonhomologous end joining / protein import into nucleus / positive regulation of fibroblast proliferation / nuclear envelope / single-stranded DNA binding / 4 iron, 4 sulfur cluster binding / DNA replication / chromosome, telomeric region / DNA-directed DNA polymerase / DNA-directed DNA polymerase activity / intracellular membrane-bounded organelle / DNA repair / nucleotide binding / DNA damage response / chromatin binding / chromatin / nucleolus / protein kinase binding / magnesium ion binding / DNA binding / zinc ion binding / nucleoplasm / membrane / nucleus / metal ion binding / cytosol 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: 4.6 Å | |||||||||
![]() | Cai SW / Zinder JC / Svetlov V / Bush MW / Nudler E / Walz T / de Lange T | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Cryo-EM structure of the human CST-Polα/primase complex in a recruitment state. 著者: Sarah W Cai / John C Zinder / Vladimir Svetlov / Martin W Bush / Evgeny Nudler / Thomas Walz / Titia de Lange / ![]() 要旨: The CST-Polα/primase complex is essential for telomere maintenance and functions to counteract resection at double-strand breaks. We report a 4.6-Å resolution cryo-EM structure of human CST- ...The CST-Polα/primase complex is essential for telomere maintenance and functions to counteract resection at double-strand breaks. We report a 4.6-Å resolution cryo-EM structure of human CST-Polα/primase, captured prior to catalysis in a recruitment state stabilized by chemical cross-linking. Our structure reveals an evolutionarily conserved interaction between the C-terminal domain of the catalytic POLA1 subunit and an N-terminal expansion in metazoan CTC1. Cross-linking mass spectrometry and negative-stain EM analysis provide insight into CST binding by the flexible POLA1 N-terminus. Finally, Coats plus syndrome disease mutations previously characterized to disrupt formation of the CST-Polα/primase complex map to protein-protein interfaces observed in the recruitment state. Together, our results shed light on the architecture and stoichiometry of the metazoan fill-in machinery. | |||||||||
履歴 |
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構造の表示
添付画像 |
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ダウンロードとリンク
-EMDBアーカイブ
マップデータ | ![]() | 95.9 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 27.8 KB 27.8 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 10.7 KB | 表示 | ![]() |
画像 | ![]() | 96.1 KB | ||
Filedesc metadata | ![]() | 9.1 KB | ||
その他 | ![]() ![]() | 80.9 MB 80.8 MB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 939 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 938.5 KB | 表示 | |
XML形式データ | ![]() | 17.3 KB | 表示 | |
CIF形式データ | ![]() | 22.9 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
関連構造データ | ![]() 7u5cMC M: このマップから作成された原子モデル C: 同じ文献を引用 ( |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
電子顕微鏡画像生データ | ![]() Data size: 869.0 Data #1: Motion corrected micrographs of CST-Pola/Primase deltaN complex in recruitment state [micrographs - single frame]) |
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リンク
EMDBのページ | ![]() ![]() |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||
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注釈 | Human CST-Pol alpha/Primase bound to ssDNA in a recruitment state. | ||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.08 Å | ||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
-ハーフマップ: #1
ファイル | emd_26346_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_26346_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 |
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試料の構成要素
+全体 : Human CST bound to DNA polymerase alpha(POLA1 delta N)-primase in...
+超分子 #1: Human CST bound to DNA polymerase alpha(POLA1 delta N)-primase in...
+分子 #1: DNA primase small subunit
+分子 #2: DNA primase large subunit
+分子 #3: DNA polymerase alpha catalytic subunit
+分子 #4: DNA polymerase alpha subunit B
+分子 #5: CST complex subunit CTC1
+分子 #6: CST complex subunit STN1
+分子 #7: CST complex subunit TEN1
+分子 #8: canonical telomeric DNA sequence
+分子 #9: ZINC ION
+分子 #10: 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 | 0.075 mg/mL |
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緩衝液 | pH: 7.5 |
グリッド | モデル: Quantifoil R1.2/1.3 / 材質: GOLD / 支持フィルム - 材質: GRAPHENE OXIDE / 前処理 - タイプ: GLOW DISCHARGE |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 278 K / 装置: FEI VITROBOT MARK IV |
詳細 | The sample was cross-linked with glutaraldehyde (GraFix) |
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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 K3 (6k x 4k) / 平均電子線量: 52.0 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: OTHER / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 2.2 µm / 最小 デフォーカス(公称値): 1.0 µm |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |