+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-13375 | |||||||||
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タイトル | Core human replisome | |||||||||
マップデータ | Consensus refinement core human replisome | |||||||||
試料 |
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キーワード | DNA replication / helicase / CMG / TIMELESS / TIPIN / CLASPIN / AND-1 / EPSILON / POLYMERASE / REPLISOME / REPLICATION | |||||||||
機能・相同性 | 機能・相同性情報 cellular response to bleomycin / DNA secondary structure binding / detection of abiotic stimulus / replication fork arrest / regulation of nuclear cell cycle DNA replication / Switching of origins to a post-replicative state / Unwinding of DNA / cell cycle phase transition / cellular response to cisplatin / DNA replication initiation ...cellular response to bleomycin / DNA secondary structure binding / detection of abiotic stimulus / replication fork arrest / regulation of nuclear cell cycle DNA replication / Switching of origins to a post-replicative state / Unwinding of DNA / cell cycle phase transition / cellular response to cisplatin / DNA replication initiation / epsilon DNA polymerase complex / DNA strand elongation involved in mitotic DNA replication / GINS complex / mitotic DNA replication preinitiation complex assembly / nuclear origin of replication recognition complex / cellular response to hydroxyurea / nucleotide-excision repair, DNA gap filling / anaphase-promoting complex binding / mitotic DNA replication / alpha DNA polymerase:primase complex / DNA replication proofreading / CMG complex / DNA replication checkpoint signaling / single-stranded DNA 3'-5' DNA exonuclease activity / DNA replication preinitiation complex / MCM complex / replication fork protection complex / regulation of phosphorylation / mitotic DNA replication checkpoint signaling / double-strand break repair via break-induced replication / mitotic DNA replication initiation / regulation of DNA-templated DNA replication initiation / mitotic intra-S DNA damage checkpoint signaling / DNA strand elongation involved in DNA replication / 加水分解酵素; エステル加水分解酵素; 5'-リン酸モノエステル産生エンドデオキシリボヌクレアーゼ / positive regulation of double-strand break repair / inner cell mass cell proliferation / activation of protein kinase activity / branching morphogenesis of an epithelial tube / DNA synthesis involved in DNA repair / cochlea development / leading strand elongation / DNA unwinding involved in DNA replication / G1/S-Specific Transcription / Apoptotic cleavage of cellular proteins / replication fork processing / nuclear replication fork / mitotic G2 DNA damage checkpoint signaling / DNA replication origin binding / positive regulation of double-strand break repair via homologous recombination / DNA replication initiation / PCNA-Dependent Long Patch Base Excision Repair / Activation of the pre-replicative complex / error-prone translesion synthesis / embryonic organ development / cellular response to interleukin-4 / Activation of ATR in response to replication stress / response to UV / base-excision repair, gap-filling / DNA helicase activity / Gap-filling DNA repair synthesis and ligation in GG-NER / ciliary basal body / DNA damage checkpoint signaling / morphogenesis of an epithelium / Assembly of the pre-replicative complex / helicase activity / Recognition of DNA damage by PCNA-containing replication complex / lung development / Termination of translesion DNA synthesis / regulation of circadian rhythm / HDR through Homologous Recombination (HRR) / Dual Incision in GG-NER / DNA-templated DNA replication / Dual incision in TC-NER / Gap-filling DNA repair synthesis and ligation in TC-NER / Orc1 removal from chromatin / circadian rhythm / G1/S transition of mitotic cell cycle / cellular response to xenobiotic stimulus / nucleosome assembly / site of double-strand break / mitotic cell cycle / single-stranded DNA binding / Processing of DNA double-strand break ends / 4 iron, 4 sulfur cluster binding / histone binding / peptidyl-serine phosphorylation / DNA helicase / DNA replication / cell population proliferation / DNA-directed DNA polymerase / chromosome, telomeric region / DNA-directed DNA polymerase activity / nuclear body / Ub-specific processing proteases / cell division / intracellular membrane-bounded organelle / DNA repair / negative regulation of DNA-templated transcription / nucleotide 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: 3.2 Å | |||||||||
データ登録者 | Jones MJ / Yeeles JTP | |||||||||
資金援助 | 英国, 1件
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引用 | ジャーナル: Nature / 年: 2021 タイトル: A conserved mechanism for regulating replisome disassembly in eukaryotes. 著者: Michael Jenkyn-Bedford / Morgan L Jones / Yasemin Baris / Karim P M Labib / Giuseppe Cannone / Joseph T P Yeeles / Tom D Deegan / 要旨: Replisome disassembly is the final step of eukaryotic DNA replication and is triggered by ubiquitylation of the CDC45-MCM-GINS (CMG) replicative helicase. Despite being driven by evolutionarily ...Replisome disassembly is the final step of eukaryotic DNA replication and is triggered by ubiquitylation of the CDC45-MCM-GINS (CMG) replicative helicase. Despite being driven by evolutionarily diverse E3 ubiquitin ligases in different eukaryotes (SCF in budding yeast, CUL2 in metazoa), replisome disassembly is governed by a common regulatory principle, in which ubiquitylation of CMG is suppressed before replication termination, to prevent replication fork collapse. Recent evidence suggests that this suppression is mediated by replication fork DNA. However, it is unknown how SCF and CUL2 discriminate terminated from elongating replisomes, to selectively ubiquitylate CMG only after termination. Here we used cryo-electron microscopy to solve high-resolution structures of budding yeast and human replisome-E3 ligase assemblies. Our structures show that the leucine-rich repeat domains of Dia2 and LRR1 are structurally distinct, but bind to a common site on CMG, including the MCM3 and MCM5 zinc-finger domains. The LRR-MCM interaction is essential for replisome disassembly and, crucially, is occluded by the excluded DNA strand at replication forks, establishing the structural basis for the suppression of CMG ubiquitylation before termination. Our results elucidate a conserved mechanism for the regulation of replisome disassembly in eukaryotes, and reveal a previously unanticipated role for DNA in preserving replisome integrity. | |||||||||
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_13375.map.gz | 259.8 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-13375-v30.xml emd-13375.xml | 55.1 KB 55.1 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_13375_fsc.xml | 15.6 KB | 表示 | FSCデータファイル |
画像 | emd_13375.png | 171.9 KB | ||
マスクデータ | emd_13375_msk_1.map | 325 MB | マスクマップ | |
Filedesc metadata | emd-13375.cif.gz | 16.3 KB | ||
その他 | emd_13375_additional_1.map.gz emd_13375_additional_2.map.gz emd_13375_half_map_1.map.gz emd_13375_half_map_2.map.gz | 177.6 MB 177.6 MB 260.6 MB 260.6 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-13375 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-13375 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_13375_validation.pdf.gz | 1 MB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_13375_full_validation.pdf.gz | 1 MB | 表示 | |
XML形式データ | emd_13375_validation.xml.gz | 23.3 KB | 表示 | |
CIF形式データ | emd_13375_validation.cif.gz | 30.9 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-13375 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-13375 | 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_13375.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 | Consensus refinement core human replisome | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.07 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_13375_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: Local resolution filtered consensus refinement core human replisome
ファイル | emd_13375_additional_1.map | ||||||||||||
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注釈 | Local resolution filtered consensus refinement core human replisome | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-追加マップ: Composite map generate using PHENIX combine focussed maps. Combining consensus...
ファイル | emd_13375_additional_2.map | ||||||||||||
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注釈 | Composite map generate using PHENIX combine_focussed_maps. Combining consensus refinement, AND1/CDC45/GINS and POLE/CDC45/GINS multibodies. | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: Consensus refinement core human replisome half map 2
ファイル | emd_13375_half_map_1.map | ||||||||||||
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注釈 | Consensus refinement core human replisome half map 2 | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: Consensus refinement core human replisome half map 1
ファイル | emd_13375_half_map_2.map | ||||||||||||
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注釈 | Consensus refinement core human replisome half map 1 | ||||||||||||
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 : Core human replisome
+超分子 #1: Core human replisome
+分子 #1: DNA replication licensing factor MCM2
+分子 #2: DNA replication licensing factor MCM3
+分子 #3: DNA replication licensing factor MCM4
+分子 #4: DNA replication licensing factor MCM5
+分子 #5: DNA replication licensing factor MCM6
+分子 #6: DNA replication licensing factor MCM7
+分子 #7: DNA polymerase epsilon subunit 2
+分子 #8: DNA polymerase epsilon catalytic subunit A
+分子 #9: Cell division control protein 45 homolog
+分子 #10: DNA replication complex GINS protein PSF1
+分子 #11: DNA replication complex GINS protein PSF2
+分子 #12: DNA replication complex GINS protein PSF3
+分子 #13: DNA replication complex GINS protein SLD5
+分子 #14: WD repeat and HMG-box DNA-binding protein 1
+分子 #15: Protein timeless homolog
+分子 #16: TIMELESS-interacting protein
+分子 #19: Claspin
+分子 #17: Leading strand DNA
+分子 #18: Lagging Strand DNA
+分子 #20: ZINC ION
+分子 #21: MAGNESIUM ION
+分子 #22: PHOSPHOAMINOPHOSPHONIC ACID-ADENYLATE ESTER
+分子 #23: SULFATE ION
-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: 7.8 |
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グリッド | モデル: Quantifoil R2/2 / 材質: COPPER / メッシュ: 400 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: CONTINUOUS / 前処理 - タイプ: GLOW DISCHARGE |
凍結 | 凍結剤: ETHANE |
-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 K2 SUMMIT (4k x 4k) 検出モード: COUNTING / 平均電子線量: 39.8 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 0.1 mm / 最大 デフォーカス(公称値): 3.5 µm / 最小 デフォーカス(公称値): 1.5 µm |
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 |
+画像解析
-原子モデル構築 1
精密化 | 空間: REAL |
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得られたモデル | PDB-7pfo: |