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データを開く
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基本情報
| 登録情報 | データベース: PDB / ID: 9q9f | |||||||||||||||||||||||||||
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| タイトル | DNA-PK bound to a 153 bp H2AX nucleosome model 1 | |||||||||||||||||||||||||||
要素 |
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キーワード | DNA BINDING PROTEIN / DNA-binding protein / NHEJ / Ku70/80 / nucleosome | |||||||||||||||||||||||||||
| 機能・相同性 | 機能・相同性情報positive regulation of platelet formation / Ku70:Ku80 complex / T cell receptor V(D)J recombination / negative regulation of t-circle formation / pro-B cell differentiation / DNA end binding / DNA-dependent protein kinase activity / small-subunit processome assembly / positive regulation of lymphocyte differentiation / histone H2AXS139 kinase activity ...positive regulation of platelet formation / Ku70:Ku80 complex / T cell receptor V(D)J recombination / negative regulation of t-circle formation / pro-B cell differentiation / DNA end binding / DNA-dependent protein kinase activity / small-subunit processome assembly / positive regulation of lymphocyte differentiation / histone H2AXS139 kinase activity / DNA-dependent protein kinase complex / DNA-dependent protein kinase-DNA ligase 4 complex / immunoglobulin V(D)J recombination / nonhomologous end joining complex / immature B cell differentiation / regulation of smooth muscle cell proliferation / cellular response to X-ray / regulation of epithelial cell proliferation / double-strand break repair via alternative nonhomologous end joining / double-strand break repair via classical nonhomologous end joining / nuclear telomere cap complex / Cytosolic sensors of pathogen-associated DNA / protein localization to site of double-strand break / XY body / telomere capping / IRF3-mediated induction of type I IFN / chromatin-protein adaptor activity / regulation of hematopoietic stem cell differentiation / regulation of telomere maintenance / recombinational repair / protein localization to chromosome, telomeric region / U3 snoRNA binding / peptidyl-threonine phosphorylation / positive regulation of neurogenesis / maturation of 5.8S rRNA / T cell lineage commitment / cellular hyperosmotic salinity response / positive regulation of double-strand break repair via nonhomologous end joining / 2-LTR circle formation / negative regulation of cGAS/STING signaling pathway / B cell lineage commitment / hematopoietic stem cell proliferation / response to ionizing radiation / telomeric repeat DNA binding / negative regulation of protein phosphorylation / DNA 3'-5' helicase / 5'-deoxyribose-5-phosphate lyase activity / hematopoietic stem cell differentiation / negative regulation of tumor necrosis factor-mediated signaling pathway / ectopic germ cell programmed cell death / ATP-dependent activity, acting on DNA / somitogenesis / telomere maintenance via telomerase / site of DNA damage / negative regulation of megakaryocyte differentiation / protein localization to CENP-A containing chromatin / Chromatin modifying enzymes / Replacement of protamines by nucleosomes in the male pronucleus / mitotic G1 DNA damage checkpoint signaling / CENP-A containing nucleosome / Packaging Of Telomere Ends / Recognition and association of DNA glycosylase with site containing an affected purine / Cleavage of the damaged purine / activation of innate immune response / Deposition of new CENPA-containing nucleosomes at the centromere / epigenetic regulation of gene expression / neurogenesis / telomere organization / positive regulation of erythrocyte differentiation / Interleukin-7 signaling / Recognition and association of DNA glycosylase with site containing an affected pyrimidine / Cleavage of the damaged pyrimidine / telomere maintenance / RNA Polymerase I Promoter Opening / DNA damage checkpoint signaling / Inhibition of DNA recombination at telomere / cyclin binding / Assembly of the ORC complex at the origin of replication / Meiotic synapsis / SUMOylation of chromatin organization proteins / Regulation of endogenous retroelements by the Human Silencing Hub (HUSH) complex / DNA-(apurinic or apyrimidinic site) lyase / positive regulation of DNA repair / protein modification process / DNA helicase activity / peptidyl-serine phosphorylation / replication fork / DNA methylation / Condensation of Prophase Chromosomes / cellular response to leukemia inhibitory factor / Chromatin modifications during the maternal to zygotic transition (MZT) / SIRT1 negatively regulates rRNA expression / HCMV Late Events / condensed nuclear chromosome / ERCC6 (CSB) and EHMT2 (G9a) positively regulate rRNA expression / male germ cell nucleus / response to gamma radiation / PRC2 methylates histones and DNA / positive regulation of translation / innate immune response in mucosa 類似検索 - 分子機能 | |||||||||||||||||||||||||||
| 生物種 | Homo sapiens (ヒト) | |||||||||||||||||||||||||||
| 手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 4.54 Å | |||||||||||||||||||||||||||
データ登録者 | Hall, C. / Chaplin, A.K. | |||||||||||||||||||||||||||
| 資金援助 | カナダ, 英国, 2件
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引用 | ジャーナル: Nat Commun / 年: 2025タイトル: Cryo-EM structures of NHEJ assemblies with nucleosomes. 著者: Chloe Hall / Philippe Frit / Antonia Kefala-Stavridi / Amandine Pelletier / Steven W Hardwick / Himani Amin / Matthew K Bilyard / Taiana Maia De Oliviera / Ammarah Tariq / Sayma Zahid / ...著者: Chloe Hall / Philippe Frit / Antonia Kefala-Stavridi / Amandine Pelletier / Steven W Hardwick / Himani Amin / Matthew K Bilyard / Taiana Maia De Oliviera / Ammarah Tariq / Sayma Zahid / Dimitri Y Chirgadze / Shankar Balasubramanian / Katheryn Meek / Virginie Ropars / Jean-Baptiste Charbonnier / Mauro Modesti / Patrick Calsou / Sébastien Britton / Tom L Blundell / Thomas Schalch / Amanda K Chaplin / ![]() 要旨: DNA double-strand breaks (DSBs) are highly deleterious lesions that can trigger cell death or carcinogenesis if unrepaired or misrepaired. In mammals, most DSBs are repaired by non-homologous end ...DNA double-strand breaks (DSBs) are highly deleterious lesions that can trigger cell death or carcinogenesis if unrepaired or misrepaired. In mammals, most DSBs are repaired by non-homologous end joining (NHEJ), which begins when Ku70/80 binds DNA ends and recruits DNA-PKcs to form the DNA-PK holoenzyme. Although recent cryo-EM studies have resolved several NHEJ assemblies, how these factors access DSBs within nucleosomes remains unclear. Here, we present cryo-EM structures of human Ku70/80 and DNA-PK bound to nucleosomes. Ku70/80 binds the DNA end and bends it away from the nucleosome core, while the Ku70 C-terminal SAP domain makes an additional, specific DNA contact. Our DNA-PK-nucleosome structure further reveals the opening of the Ku80 vWA domain, and we show that non-hydrolysable ATP promotes synapsis by stabilising the Ku80-mediated DNA-PK dimer. These structures reveal a model for DSB recognition on nucleosomal DNA and provide insights relevant to targeting NHEJ in cancer therapy. | |||||||||||||||||||||||||||
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構造の表示
| 構造ビューア | 分子: Molmil Jmol/JSmol |
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ダウンロードとリンク
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ダウンロード
| PDBx/mmCIF形式 | 9q9f.cif.gz | 1 MB | 表示 | PDBx/mmCIF形式 |
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| PDB形式 | pdb9q9f.ent.gz | 表示 | PDB形式 | |
| PDBx/mmJSON形式 | 9q9f.json.gz | ツリー表示 | PDBx/mmJSON形式 | |
| その他 | その他のダウンロード |
-検証レポート
| アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/q9/9q9f ftp://data.pdbj.org/pub/pdb/validation_reports/q9/9q9f | HTTPS FTP |
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-関連構造データ
| 関連構造データ | ![]() 52958MC ![]() 9igwC ![]() 9igxC ![]() 9q80C ![]() 9q8xC ![]() 9qcrC ![]() 9qcsC ![]() 9qmsC M: このデータのモデリングに利用したマップデータ 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 F&H 検索 |
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リンク
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集合体
| 登録構造単位 | ![]()
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要素
-X-ray repair cross-complementing protein ... , 2種, 2分子 KL
| #1: タンパク質 | 分子量: 69945.039 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: XRCC6, G22P1発現宿主: ![]() 参照: UniProt: P12956, 加水分解酵素; 酸無水物に作用; 酸無水物に作用・細胞または細胞小器官の運動に関与, 付加脱離酵素(リアーゼ); 炭素- ...参照: UniProt: P12956, 加水分解酵素; 酸無水物に作用; 酸無水物に作用・細胞または細胞小器官の運動に関与, 付加脱離酵素(リアーゼ); 炭素-酸素リアーゼ類; その他の炭素-酸素リアーゼ |
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| #2: タンパク質 | 分子量: 82812.438 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: XRCC5, G22P2発現宿主: ![]() 参照: UniProt: P13010, 加水分解酵素; 酸無水物に作用; 酸無水物に作用・細胞または細胞小器官の運動に関与 |
-DNA鎖 , 2種, 2分子 IJ
| #3: DNA鎖 | 分子量: 46998.945 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 発現宿主: ![]() |
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| #4: DNA鎖 | 分子量: 47457.234 Da / 分子数: 1 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 発現宿主: ![]() |
-タンパク質 , 5種, 9分子 OCGDHAEBF
| #5: タンパク質 | 分子量: 469673.219 Da / 分子数: 1 / 由来タイプ: 天然 / 由来: (天然) Homo sapiens (ヒト)参照: UniProt: P78527, non-specific serine/threonine protein kinase | ||||||
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| #6: タンパク質 | 分子量: 15172.605 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: H2AX, H2AFX / 発現宿主: ![]() #7: タンパク質 | 分子量: 13935.239 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: H2BC11, H2BFR, HIST1H2BJ / 発現宿主: ![]() #8: タンパク質 | 分子量: 15437.167 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト)遺伝子: HIST1H3A, H3FA, HIST1H3B, H3FL, HIST1H3C, H3FC, HIST1H3D, H3FB, HIST1H3E, H3FD, HIST1H3F, H3FI, HIST1H3G, H3FH, HIST1H3H, H3FK, HIST1H3I, H3FF, HIST1H3J, H3FJ 発現宿主: ![]() #9: タンパク質 | 分子量: 11394.426 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト)遺伝子: H4C1, H4/A, H4FA, HIST1H4A, H4C2, H4/I, H4FI, HIST1H4B, H4C3, H4/G, H4FG, HIST1H4C, H4C4, H4/B, H4FB, HIST1H4D, H4C5, H4/J, H4FJ, HIST1H4E, H4C6, H4/C, H4FC, HIST1H4F, H4C8, H4/H, H4FH, ...遺伝子: H4C1, H4/A, H4FA, HIST1H4A, H4C2, H4/I, H4FI, HIST1H4B, H4C3, H4/G, H4FG, HIST1H4C, H4C4, H4/B, H4FB, HIST1H4D, H4C5, H4/J, H4FJ, HIST1H4E, H4C6, H4/C, H4FC, HIST1H4F, H4C8, H4/H, H4FH, HIST1H4H, H4C9, H4/M, H4FM, HIST1H4I, H4C11, H4/E, H4FE, HIST1H4J, H4C12, H4/D, H4FD, HIST1H4K, H4C13, H4/K, H4FK, HIST1H4L, H4C14, H4/N, H4F2, H4FN, HIST2H4, HIST2H4A, H4C15, H4/O, H4FO, HIST2H4B, H4C16, H4-16, HIST4H4 発現宿主: ![]() |
-詳細
| Has protein modification | N |
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-実験情報
-実験
| 実験 | 手法: 電子顕微鏡法 |
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| EM実験 | 試料の集合状態: PARTICLE / 3次元再構成法: 単粒子再構成法 |
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試料調製
| 構成要素 | 名称: DNA-PK bound to a 153 bp H2AX nucleosome model 1 / タイプ: COMPLEX / Entity ID: all / 由来: RECOMBINANT | |||||||||||||||||||||||||||||||||||
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| 分子量 | 値: 0.829 MDa / 実験値: YES | |||||||||||||||||||||||||||||||||||
| 由来(天然) | 生物種: Homo sapiens (ヒト) | |||||||||||||||||||||||||||||||||||
| 由来(組換発現) | 生物種: ![]() | |||||||||||||||||||||||||||||||||||
| 緩衝液 | pH: 7.6 | |||||||||||||||||||||||||||||||||||
| 緩衝液成分 |
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| 試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | |||||||||||||||||||||||||||||||||||
| 試料支持 | 詳細: 30mA / グリッドの材料: COPPER / グリッドのサイズ: 300 divisions/in. / グリッドのタイプ: Quantifoil R1.2/1.3 | |||||||||||||||||||||||||||||||||||
| 急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 277 K |
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電子顕微鏡撮影
| 実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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| 顕微鏡 | モデル: TFS KRIOS |
| 電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: FLOOD BEAM |
| 電子レンズ | モード: BRIGHT FIELD / 倍率(公称値): 105000 X / 最大 デフォーカス(公称値): 2200 nm / 最小 デフォーカス(公称値): 700 nm / Cs: 2.7 mm / アライメント法: ZEMLIN TABLEAU |
| 試料ホルダ | 凍結剤: NITROGEN 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
| 撮影 | 平均露光時間: 2.2 sec. / 電子線照射量: 48.59 e/Å2 / フィルム・検出器のモデル: GATAN K3 (6k x 4k) |
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解析
| EMソフトウェア |
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| CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
| 3次元再構成 | 解像度: 4.54 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 14174 / 対称性のタイプ: POINT | ||||||||||||||||||||||||
| 精密化 | 最高解像度: 4.54 Å 立体化学のターゲット値: REAL-SPACE (WEIGHTED MAP SUM AT ATOM CENTERS) | ||||||||||||||||||||||||
| 拘束条件 |
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コントローラー
万見について




Homo sapiens (ヒト)
カナダ,
英国, 2件
引用
















PDBj



















































FIELD EMISSION GUN