+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-23510 | |||||||||||||||||||||
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タイトル | NHEJ Long-range synaptic complex | |||||||||||||||||||||
マップデータ | Pre-synaptic complex | |||||||||||||||||||||
試料 |
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キーワード | NHEJ / DNA BINDING PROTEIN / DNA BINDING PROTEIN-DNA complex | |||||||||||||||||||||
機能・相同性 | 機能・相同性情報 DNA ligation involved in DNA recombination / T cell receptor V(D)J recombination / FHA domain binding / positive regulation of chromosome organization / positive regulation of ligase activity / pro-B cell differentiation / DNA ligase IV complex / positive regulation of lymphocyte differentiation / DNA ligation involved in DNA repair / small-subunit processome assembly ...DNA ligation involved in DNA recombination / T cell receptor V(D)J recombination / FHA domain binding / positive regulation of chromosome organization / positive regulation of ligase activity / pro-B cell differentiation / DNA ligase IV complex / positive regulation of lymphocyte differentiation / DNA ligation involved in DNA repair / small-subunit processome assembly / DNA ligase activity / Ku70:Ku80 complex / DN2 thymocyte differentiation / DNA-dependent protein kinase complex / immunoglobulin V(D)J recombination / negative regulation of t-circle formation / DNA end binding / DNA ligase (ATP) / DNA-dependent protein kinase-DNA ligase 4 complex / MHC class II antigen presentation / nonhomologous end joining complex / DNA ligase (ATP) activity / cellular response to X-ray / regulation of smooth muscle cell proliferation / single strand break repair / Cytosolic sensors of pathogen-associated DNA / nucleotide-excision repair, DNA gap filling / Neutrophil degranulation / DNA ligation / V(D)J recombination / IRF3-mediated induction of type I IFN / nuclear telomere cap complex / double-strand break repair via classical nonhomologous end joining / isotype switching / protein localization to site of double-strand break / positive regulation of catalytic activity / entry into host cell by a symbiont-containing vacuole / U3 snoRNA binding / recombinational repair / regulation of telomere maintenance / protein localization to chromosome, telomeric region / cellular response to fatty acid / positive regulation of neurogenesis / cellular hyperosmotic salinity response / hematopoietic stem cell proliferation / response to ionizing radiation / cellular response to lithium ion / 加水分解酵素; プロテアーゼ; ペプチド結合加水分解酵素; アスパラギン酸プロテアーゼ / DNA biosynthetic process / telomeric DNA binding / 2-LTR circle formation / ligase activity / : / site of DNA damage / somatic stem cell population maintenance / protein autoprocessing / 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 / response to X-ray / T cell differentiation / 5'-deoxyribose-5-phosphate lyase activity / hematopoietic stem cell differentiation / positive regulation of protein kinase activity / chromosome organization / ATP-dependent activity, acting on DNA / SUMOylation of DNA damage response and repair proteins / DNA polymerase binding / condensed chromosome / activation of innate immune response / transport vesicle / enzyme activator activity / positive regulation of telomere maintenance via telomerase / DNA helicase activity / telomere maintenance / cyclin binding / B cell differentiation / neurogenesis / protein-DNA complex / stem cell proliferation / response to gamma radiation / cellular response to leukemia inhibitory factor / central nervous system development / small-subunit processome / cellular response to ionizing radiation / Nonhomologous End-Joining (NHEJ) / 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 / cellular response to gamma radiation / fibrillar center / protein processing / double-strand break repair via nonhomologous end joining / establishment of integrated proviral latency / positive regulation of fibroblast proliferation / double-strand break repair / site of double-strand break / T cell differentiation in thymus / double-stranded DNA binding / fibroblast proliferation / scaffold protein binding / secretory granule lumen / DNA recombination / in utero embryonic development 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: 4.6 Å | |||||||||||||||||||||
データ登録者 | He Y / Chen S | |||||||||||||||||||||
資金援助 | 米国, 6件
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引用 | ジャーナル: Nature / 年: 2021 タイトル: Structural basis of long-range to short-range synaptic transition in NHEJ. 著者: Siyu Chen / Linda Lee / Tasmin Naila / Susan Fishbain / Annie Wang / Alan E Tomkinson / Susan P Lees-Miller / Yuan He / 要旨: DNA double-strand breaks (DSBs) are a highly cytotoxic form of DNA damage and the incorrect repair of DSBs is linked to carcinogenesis. The conserved error-prone non-homologous end joining (NHEJ) ...DNA double-strand breaks (DSBs) are a highly cytotoxic form of DNA damage and the incorrect repair of DSBs is linked to carcinogenesis. The conserved error-prone non-homologous end joining (NHEJ) pathway has a key role in determining the effects of DSB-inducing agents that are used to treat cancer as well as the generation of the diversity in antibodies and T cell receptors. Here we applied single-particle cryo-electron microscopy to visualize two key DNA-protein complexes that are formed by human NHEJ factors. The Ku70/80 heterodimer (Ku), the catalytic subunit of the DNA-dependent protein kinase (DNA-PKcs), DNA ligase IV (LigIV), XRCC4 and XLF form a long-range synaptic complex, in which the DNA ends are held approximately 115 Å apart. Two DNA end-bound subcomplexes comprising Ku and DNA-PKcs are linked by interactions between the DNA-PKcs subunits and a scaffold comprising LigIV, XRCC4, XLF, XRCC4 and LigIV. The relative orientation of the DNA-PKcs molecules suggests a mechanism for autophosphorylation in trans, which leads to the dissociation of DNA-PKcs and the transition into the short-range synaptic complex. Within this complex, the Ku-bound DNA ends are aligned for processing and ligation by the XLF-anchored scaffold, and a single catalytic domain of LigIV is stably associated with a nick between the two Ku molecules, which suggests that the joining of both strands of a DSB involves both LigIV molecules. | |||||||||||||||||||||
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_23510.map.gz | 81.9 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-23510-v30.xml emd-23510.xml | 33 KB 33 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_23510_fsc.xml | 15.4 KB | 表示 | FSCデータファイル |
画像 | emd_23510.png | 145.8 KB | ||
Filedesc metadata | emd-23510.cif.gz | 11.7 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-23510 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-23510 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_23510_validation.pdf.gz | 481.3 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_23510_full_validation.pdf.gz | 480.9 KB | 表示 | |
XML形式データ | emd_23510_validation.xml.gz | 13.6 KB | 表示 | |
CIF形式データ | emd_23510_validation.cif.gz | 18.6 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23510 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23510 | 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_23510.map.gz / 形式: CCP4 / 大きさ: 93 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Pre-synaptic complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.1 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 : Long-range synaptic Complex of NHEJ
+超分子 #1: Long-range synaptic Complex of NHEJ
+分子 #1: X-ray repair cross-complementing protein 6
+分子 #2: X-ray repair cross-complementing protein 5
+分子 #3: DNA-dependent protein kinase catalytic subunit
+分子 #4: Unknown peptide
+分子 #7: DNA repair protein XRCC4
+分子 #8: Non-homologous end-joining factor 1
+分子 #9: DNA ligase 4
+分子 #5: DNA (31-MER)
+分子 #6: DNA (30-MER)
+分子 #10: ADENOSINE-5'-DIPHOSPHATE
-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: 7.9 構成要素:
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グリッド | モデル: Quantifoil R3.5/1 / 材質: COPPER / メッシュ: 400 / 支持フィルム - 材質: GRAPHENE OXIDE / 支持フィルム - トポロジー: CONTINUOUS / 支持フィルム - Film thickness: 200 / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 時間: 10 sec. / 前処理 - 雰囲気: AIR / 前処理 - 気圧: 101.325 kPa | |||||||||||||||||||||
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 | JEOL 3200FS |
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撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: COUNTING / デジタル化 - サイズ - 横: 3710 pixel / デジタル化 - サイズ - 縦: 3838 pixel / デジタル化 - 画像ごとのフレーム数: 1-30 / 撮影したグリッド数: 4 / 実像数: 17114 / 平均露光時間: 0.3 sec. / 平均電子線量: 76.5 e/Å2 |
電子線 | 加速電圧: 200 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | C2レンズ絞り径: 100.0 µm / 照射モード: FLOOD BEAM / 撮影モード: DARK FIELD / Cs: 2.7 mm / 最大 デフォーカス(公称値): 4.0 µm / 最小 デフォーカス(公称値): 2.0 µm / 倍率(公称値): 30000 |
試料ステージ | 試料ホルダーモデル: JEOL / ホルダー冷却材: NITROGEN |
+画像解析
-原子モデル構築 1
初期モデル |
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精密化 | 空間: REAL / プロトコル: FLEXIBLE FIT / 当てはまり具合の基準: Correlation coefficient | ||||||||||||||||
得られたモデル | PDB-7lt3: |