+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-23755 | |||||||||||||||
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タイトル | CryoEM map of the structure of the S. cerevisiae origin recognition complex bound to the replication initiator Cdc6 and the ARS1 origin DNA. | |||||||||||||||
マップデータ | ||||||||||||||||
試料 |
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機能・相同性 | 機能・相同性情報 CDK-mediated phosphorylation and removal of Cdc6 / CDC6 association with the ORC:origin complex / Assembly of the ORC complex at the origin of replication / Orc1 removal from chromatin / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / nuclear origin of replication recognition complex / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / Activation of the pre-replicative complex / nuclear pre-replicative complex ...CDK-mediated phosphorylation and removal of Cdc6 / CDC6 association with the ORC:origin complex / Assembly of the ORC complex at the origin of replication / Orc1 removal from chromatin / Cul8-RING ubiquitin ligase complex / maintenance of rDNA / nuclear origin of replication recognition complex / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / Activation of the pre-replicative complex / nuclear pre-replicative complex / Activation of ATR in response to replication stress / DNA replication preinitiation complex / mitotic DNA replication checkpoint signaling / silent mating-type cassette heterochromatin formation / regulation of DNA-templated DNA replication initiation / DNA replication origin binding / regulation of DNA replication / subtelomeric heterochromatin formation / DNA replication initiation / nucleosome binding / G1/S transition of mitotic cell cycle / 染色体 / chromosome, telomeric region / 細胞分裂 / GTPase activity / chromatin binding / GTP binding / ATP hydrolysis activity / 核質 / ATP binding / metal ion binding / 細胞核 / 細胞質 類似検索 - 分子機能 | |||||||||||||||
生物種 | Saccharomyces cerevisiae (パン酵母) | |||||||||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.6 Å | |||||||||||||||
データ登録者 | Feng X / Li H | |||||||||||||||
資金援助 | 米国, 4件
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引用 | ジャーナル: Nat Commun / 年: 2021 タイトル: The structure of ORC-Cdc6 on an origin DNA reveals the mechanism of ORC activation by the replication initiator Cdc6. 著者: Xiang Feng / Yasunori Noguchi / Marta Barbon / Bruce Stillman / Christian Speck / Huilin Li / 要旨: The Origin Recognition Complex (ORC) binds to sites in chromosomes to specify the location of origins of DNA replication. The S. cerevisiae ORC binds to specific DNA sequences throughout the cell ...The Origin Recognition Complex (ORC) binds to sites in chromosomes to specify the location of origins of DNA replication. The S. cerevisiae ORC binds to specific DNA sequences throughout the cell cycle but becomes active only when it binds to the replication initiator Cdc6. It has been unclear at the molecular level how Cdc6 activates ORC, converting it to an active recruiter of the Mcm2-7 hexamer, the core of the replicative helicase. Here we report the cryo-EM structure at 3.3 Å resolution of the yeast ORC-Cdc6 bound to an 85-bp ARS1 origin DNA. The structure reveals that Cdc6 contributes to origin DNA recognition via its winged helix domain (WHD) and its initiator-specific motif. Cdc6 binding rearranges a short α-helix in the Orc1 AAA+ domain and the Orc2 WHD, leading to the activation of the Cdc6 ATPase and the formation of the three sites for the recruitment of Mcm2-7, none of which are present in ORC alone. The results illuminate the molecular mechanism of a critical biochemical step in the licensing of eukaryotic replication origins. | |||||||||||||||
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_23755.map.gz | 12.4 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-23755-v30.xml emd-23755.xml | 30.6 KB 30.6 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_23755_fsc.xml | 10 KB | 表示 | FSCデータファイル |
画像 | emd_23755.png | 95.9 KB | ||
その他 | emd_23755_half_map_1.map.gz emd_23755_half_map_2.map.gz | 65.4 MB 65.3 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-23755 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-23755 | 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_23755.map.gz / 形式: CCP4 / 大きさ: 83.7 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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ボクセルのサイズ | X=Y=Z: 0.828 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
-ハーフマップ: #2
ファイル | emd_23755_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: #1
ファイル | emd_23755_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 : Complex of origin replication complex with Cdc6 and ARS1 origin DNA
+超分子 #1: Complex of origin replication complex with Cdc6 and ARS1 origin DNA
+分子 #1: Origin recognition complex subunit 1
+分子 #2: Origin recognition complex subunit 2
+分子 #3: Origin recognition complex subunit 3
+分子 #4: Origin recognition complex subunit 4
+分子 #5: Origin recognition complex subunit 5
+分子 #6: Origin recognition complex subunit 6
+分子 #9: Cell division control protein 6
+分子 #7: DNA (85-MER)
+分子 #8: DNA (85-MER)
+分子 #10: MAGNESIUM ION
+分子 #11: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER
-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: 7.5 構成要素:
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グリッド | モデル: Quantifoil R1.2/1.3 / 材質: COPPER / メッシュ: 300 / 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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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 FIELDBright-field microscopy |
撮影 | フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 平均電子線量: 76.0 e/Å2 |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |