+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-4465 | |||||||||
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タイトル | Cryo-EM structure of the APC/C-Cdc20-Cdk2-cyclinA2-Cks2 complex, the D1 box class | |||||||||
マップデータ | unsharpened map | |||||||||
試料 |
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キーワード | spindle assembly checkpoint / anaphase-promoting complex / cyclin / ubiquitination / CELL CYCLE | |||||||||
機能・相同性 | 機能・相同性情報 metaphase/anaphase transition of cell cycle / metaphase/anaphase transition of meiosis I / Inhibition of the proteolytic activity of APC/C required for the onset of anaphase by mitotic spindle checkpoint components / mitotic checkpoint complex / positive regulation of anaphase-promoting complex-dependent catabolic process / regulation of meiotic nuclear division / positive regulation of synapse maturation / Conversion from APC/C:Cdc20 to APC/C:Cdh1 in late anaphase / regulation of mitotic cell cycle spindle assembly checkpoint / protein branched polyubiquitination ...metaphase/anaphase transition of cell cycle / metaphase/anaphase transition of meiosis I / Inhibition of the proteolytic activity of APC/C required for the onset of anaphase by mitotic spindle checkpoint components / mitotic checkpoint complex / positive regulation of anaphase-promoting complex-dependent catabolic process / regulation of meiotic nuclear division / positive regulation of synapse maturation / Conversion from APC/C:Cdc20 to APC/C:Cdh1 in late anaphase / regulation of mitotic cell cycle spindle assembly checkpoint / protein branched polyubiquitination / Inactivation of APC/C via direct inhibition of the APC/C complex / APC/C:Cdc20 mediated degradation of mitotic proteins / Phosphorylation of proteins involved in the G2/M transition by Cyclin A:Cdc2 complexes / Phosphorylation of Emi1 / cyclin A2-CDK1 complex / anaphase-promoting complex / Aberrant regulation of mitotic exit in cancer due to RB1 defects / cell cycle G1/S phase transition / regulation of meiotic cell cycle / anaphase-promoting complex-dependent catabolic process / metaphase/anaphase transition of mitotic cell cycle / positive regulation of synaptic plasticity / cellular response to luteinizing hormone stimulus / regulation of exit from mitosis / anaphase-promoting complex binding / Phosphorylation of the APC/C / mitotic cell cycle phase transition / cellular response to leptin stimulus / positive regulation of mitotic metaphase/anaphase transition / positive regulation of ubiquitin protein ligase activity / ubiquitin ligase activator activity / Transcription of E2F targets under negative control by p107 (RBL1) and p130 (RBL2) in complex with HDAC1 / male pronucleus / protein K11-linked ubiquitination / female pronucleus / enzyme-substrate adaptor activity / positive regulation of dendrite morphogenesis / cellular response to cocaine / regulation of mitotic metaphase/anaphase transition / response to glucagon / cyclin-dependent protein serine/threonine kinase regulator activity / ubiquitin-ubiquitin ligase activity / mitotic sister chromatid cohesion / mitotic metaphase chromosome alignment / positive regulation of DNA biosynthetic process / cellular response to insulin-like growth factor stimulus / cochlea development / cyclin A2-CDK2 complex / G2 Phase / mitotic spindle assembly checkpoint signaling / p53-Dependent G1 DNA Damage Response / cellular response to platelet-derived growth factor stimulus / regulation of DNA replication / Regulation of APC/C activators between G1/S and early anaphase / cullin family protein binding / G0 and Early G1 / Telomere Extension By Telomerase / Transcriptional Regulation by VENTX / mitotic spindle assembly / cellular response to nitric oxide / ubiquitin-like ligase-substrate adaptor activity / animal organ regeneration / positive regulation of axon extension / TP53 Regulates Transcription of Genes Involved in G1 Cell Cycle Arrest / protein K48-linked ubiquitination / heterochromatin / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / Cyclin A/B1/B2 associated events during G2/M transition / cyclin-dependent protein kinase holoenzyme complex / Cyclin A:Cdk2-associated events at S phase entry / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / regulation of mitotic cell cycle / Resolution of Sister Chromatid Cohesion / APC/C:Cdc20 mediated degradation of Cyclin B / post-translational protein modification / APC-Cdc20 mediated degradation of Nek2A / nuclear periphery / Autodegradation of Cdh1 by Cdh1:APC/C / APC/C:Cdc20 mediated degradation of Securin / SCF-beta-TrCP mediated degradation of Emi1 / cellular response to estradiol stimulus / Assembly of the pre-replicative complex / RHO GTPases Activate Formins / Cdc20:Phospho-APC/C mediated degradation of Cyclin A / brain development / APC/C:Cdh1 mediated degradation of Cdc20 and other APC/C:Cdh1 targeted proteins in late mitosis/early G1 / DNA Damage/Telomere Stress Induced Senescence / CDK-mediated phosphorylation and removal of Cdc6 / mitotic spindle / kinetochore / SCF(Skp2)-mediated degradation of p27/p21 / spindle pole / spindle / Orc1 removal from chromatin / ubiquitin-protein transferase activity / G1/S transition of mitotic cell cycle / Separation of Sister Chromatids / G2/M transition of mitotic cell cycle / microtubule cytoskeleton 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 | Zhang S / Barford D | |||||||||
資金援助 | 英国, 2件
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引用 | ジャーナル: Nat Commun / 年: 2019 タイトル: Cyclin A2 degradation during the spindle assembly checkpoint requires multiple binding modes to the APC/C. 著者: Suyang Zhang / Thomas Tischer / David Barford / 要旨: The anaphase-promoting complex/cyclosome (APC/C) orchestrates cell cycle progression by controlling the temporal degradation of specific cell cycle regulators. Although cyclin A2 and cyclin B1 are ...The anaphase-promoting complex/cyclosome (APC/C) orchestrates cell cycle progression by controlling the temporal degradation of specific cell cycle regulators. Although cyclin A2 and cyclin B1 are both targeted for degradation by the APC/C, during the spindle assembly checkpoint (SAC), the mitotic checkpoint complex (MCC) represses APC/C's activity towards cyclin B1, but not cyclin A2. Through structural, biochemical and in vivo analysis, we identify a non-canonical D box (D2) that is critical for cyclin A2 ubiquitination in vitro and degradation in vivo. During the SAC, cyclin A2 is ubiquitinated by the repressed APC/C-MCC, mediated by the cooperative engagement of its KEN and D2 boxes, ABBA motif, and the cofactor Cks. Once the SAC is satisfied, cyclin A2 binds APC/C-Cdc20 through two mutually exclusive binding modes, resulting in differential ubiquitination efficiency. Our findings reveal that a single substrate can engage an E3 ligase through multiple binding modes, affecting its degradation timing and efficiency. | |||||||||
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_4465.map.gz | 213.7 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-4465-v30.xml emd-4465.xml | 43.8 KB 43.8 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_4465_fsc.xml | 14.2 KB | 表示 | FSCデータファイル |
画像 | emd_4465.png | 160.8 KB | ||
Filedesc metadata | emd-4465.cif.gz | 12 KB | ||
その他 | emd_4465_additional.map.gz emd_4465_half_map_1.map.gz emd_4465_half_map_2.map.gz | 10.2 MB 194.7 MB 194.7 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-4465 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-4465 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_4465_validation.pdf.gz | 429.8 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_4465_full_validation.pdf.gz | 428.9 KB | 表示 | |
XML形式データ | emd_4465_validation.xml.gz | 19.8 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4465 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4465 | 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_4465.map.gz / 形式: CCP4 / 大きさ: 244.1 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | unsharpened map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.047 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
-追加マップ: sharpened map with B factor -30 and lowpass filtered to 4A
ファイル | emd_4465_additional.map | ||||||||||||
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注釈 | sharpened map with B factor -30 and lowpass filtered to 4A | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: halfmap1
ファイル | emd_4465_half_map_1.map | ||||||||||||
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注釈 | halfmap1 | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: halfmap2
ファイル | emd_4465_half_map_2.map | ||||||||||||
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注釈 | halfmap2 | ||||||||||||
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 : Cryo-EM structure of the APC/C-Cdc20-Cdk2-cyclinA2-Cks2 complex, ...
+超分子 #1: Cryo-EM structure of the APC/C-Cdc20-Cdk2-cyclinA2-Cks2 complex, ...
+超分子 #2: Cryo-EM structure of the APC/C-Cdc20-Cdk2-cyclinA2-Cks2 complex, ...
+超分子 #3: Cryo-EM structure of the APC/C-Cdc20-Cdk2-cyclinA2-Cks2 complex, ...
+分子 #1: Cell division cycle protein 20 homolog
+分子 #2: Cyclin-A2
+分子 #3: Anaphase-promoting complex subunit 10
+分子 #4: Anaphase-promoting complex subunit 15
+分子 #5: Anaphase-promoting complex subunit 1,Anaphase-promoting complex s...
+分子 #6: Anaphase-promoting complex subunit 2
+分子 #7: Anaphase-promoting complex subunit 4
+分子 #8: Anaphase-promoting complex subunit 5
+分子 #9: Cell division cycle protein 16 homolog
+分子 #10: Anaphase-promoting complex subunit 11
+分子 #11: Anaphase-promoting complex subunit CDC26
+分子 #12: Anaphase-promoting complex subunit 13
+分子 #13: Anaphase-promoting complex subunit 16
+分子 #14: Cell division cycle protein 27 homolog
+分子 #15: Anaphase-promoting complex subunit 7
+分子 #16: Cell division cycle protein 23 homolog
-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
濃度 | 0.15 mg/mL |
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緩衝液 | pH: 8 / 詳細: 20mM Hepes, 150mM NaCl, 0.5mM TCEP |
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) 平均電子線量: 28.0 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 |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |