+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-32252 | ||||||||||||
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タイトル | Cryo-EM structure of human cohesin-CTCF-DNA complex | ||||||||||||
マップデータ | |||||||||||||
試料 |
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キーワード | Cohesin / NIPBL / CTCF / DNA / chromosome folding / topologically associating domain / chromatin loops / DNA loop extrusion / sister chromatid cohesion / complex / ATPase / HEAT repeat protein / DNA BINDING PROTEIN / DNA BINDING PROTEIN-DNA complex | ||||||||||||
機能・相同性 | 機能・相同性情報 eye morphogenesis / cohesin loader activity / external genitalia morphogenesis / gallbladder development / SMC loading complex / Scc2-Scc4 cohesin loading complex / ear morphogenesis / mitotic cohesin loading / regulation of hair cycle / chromatin loop anchoring activity ...eye morphogenesis / cohesin loader activity / external genitalia morphogenesis / gallbladder development / SMC loading complex / Scc2-Scc4 cohesin loading complex / ear morphogenesis / mitotic cohesin loading / regulation of hair cycle / chromatin loop anchoring activity / chromatin insulator sequence binding / response to DNA damage checkpoint signaling / maintenance of mitotic sister chromatid cohesion / forelimb morphogenesis / negative regulation of mitotic metaphase/anaphase transition / embryonic viscerocranium morphogenesis / positive regulation of sister chromatid cohesion / Cohesin Loading onto Chromatin / meiotic cohesin complex / Establishment of Sister Chromatid Cohesion / establishment of meiotic sister chromatid cohesion / cohesin complex / mitotic cohesin complex / uterus morphogenesis / embryonic digestive tract morphogenesis / regulation of centromeric sister chromatid cohesion / negative regulation of G2/M transition of mitotic cell cycle / establishment of protein localization to chromatin / regulation of developmental growth / negative regulation of glial cell apoptotic process / genomic imprinting / cellular response to X-ray / positive regulation of neuron migration / integrator complex / chromo shadow domain binding / lateral element / replication-born double-strand break repair via sister chromatid exchange / mediator complex binding / protein localization to chromosome, centromeric region / establishment of mitotic sister chromatid cohesion / digestive tract development / positive regulation of multicellular organism growth / metanephros development / positive regulation of ossification / chromatin looping / embryonic forelimb morphogenesis / reciprocal meiotic recombination / face morphogenesis / sister chromatid cohesion / negative regulation of interleukin-1 beta production / microtubule motor activity / lncRNA binding / negative regulation of gene expression via chromosomal CpG island methylation / mitotic sister chromatid cohesion / stem cell population maintenance / dynein complex binding / fat cell differentiation / mitotic spindle pole / beta-tubulin binding / regulation of DNA replication / outflow tract morphogenesis / mitotic sister chromatid segregation / somatic stem cell population maintenance / regulation of embryonic development / positive regulation of interleukin-10 production / negative regulation of tumor necrosis factor production / chromosome, centromeric region / developmental growth / mitotic spindle assembly / heart morphogenesis / SUMOylation of DNA damage response and repair proteins / condensed chromosome / protein localization to chromatin / Resolution of Sister Chromatid Cohesion / Meiotic synapsis / epigenetic regulation of gene expression / condensed nuclear chromosome / male germ cell nucleus / meiotic cell cycle / transcription coregulator binding / chromosome segregation / promoter-specific chromatin binding / sensory perception of sound / brain development / response to radiation / kinetochore / DNA-binding transcription repressor activity, RNA polymerase II-specific / histone deacetylase binding / cognition / spindle pole / nuclear matrix / Activation of anterior HOX genes in hindbrain development during early embryogenesis / Separation of Sister Chromatids / transcription corepressor activity / sequence-specific double-stranded DNA binding / protein localization / double-strand break repair / mitotic cell cycle / chromosome / double-stranded DNA 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: 6.5 Å | ||||||||||||
データ登録者 | Shi ZB / Bai XC | ||||||||||||
資金援助 | 米国, 3件
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引用 | ジャーナル: Mol Cell / 年: 2023 タイトル: CTCF and R-loops are boundaries of cohesin-mediated DNA looping. 著者: Hongshan Zhang / Zhubing Shi / Edward J Banigan / Yoori Kim / Hongtao Yu / Xiao-Chen Bai / Ilya J Finkelstein / 要旨: Cohesin and CCCTC-binding factor (CTCF) are key regulatory proteins of three-dimensional (3D) genome organization. Cohesin extrudes DNA loops that are anchored by CTCF in a polar orientation. Here, ...Cohesin and CCCTC-binding factor (CTCF) are key regulatory proteins of three-dimensional (3D) genome organization. Cohesin extrudes DNA loops that are anchored by CTCF in a polar orientation. Here, we present direct evidence that CTCF binding polarity controls cohesin-mediated DNA looping. Using single-molecule imaging, we demonstrate that a critical N-terminal motif of CTCF blocks cohesin translocation and DNA looping. The cryo-EM structure of the cohesin-CTCF complex reveals that this CTCF motif ahead of zinc fingers can only reach its binding site on the STAG1 cohesin subunit when the N terminus of CTCF faces cohesin. Remarkably, a C-terminally oriented CTCF accelerates DNA compaction by cohesin. DNA-bound Cas9 and Cas12a ribonucleoproteins are also polar cohesin barriers, indicating that stalling may be intrinsic to cohesin itself. Finally, we show that RNA-DNA hybrids (R-loops) block cohesin-mediated DNA compaction in vitro and are enriched with cohesin subunits in vivo, likely forming TAD boundaries. | ||||||||||||
履歴 |
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-構造の表示
添付画像 |
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-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_32252.map.gz | 104.5 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-32252-v30.xml emd-32252.xml | 29.9 KB 29.9 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_32252_fsc.xml | 12.8 KB | 表示 | FSCデータファイル |
画像 | emd_32252.png | 63.1 KB | ||
Filedesc metadata | emd-32252.cif.gz | 11.1 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-32252 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-32252 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_32252_validation.pdf.gz | 402.9 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_32252_full_validation.pdf.gz | 402.5 KB | 表示 | |
XML形式データ | emd_32252_validation.xml.gz | 12.9 KB | 表示 | |
CIF形式データ | emd_32252_validation.cif.gz | 17.6 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-32252 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-32252 | HTTPS FTP |
-関連構造データ
関連構造データ | 7w1mMC M: このマップから作成された原子モデル C: 同じ文献を引用 (文献) |
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類似構造データ | 類似検索 - 機能・相同性F&H 検索 |
-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_32252.map.gz / 形式: CCP4 / 大きさ: 178 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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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: 1.44 Å | ||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
-試料の構成要素
+全体 : Human cohesin-NIPBL-CTCF-DNA complex
+超分子 #1: Human cohesin-NIPBL-CTCF-DNA complex
+超分子 #2: Human cohesin-NIPBL-CTCF
+超分子 #3: DNA
+分子 #1: Structural maintenance of chromosomes protein 1A
+分子 #2: Structural maintenance of chromosomes protein 3
+分子 #3: Double-strand-break repair protein rad21 homolog
+分子 #4: Cohesin subunit SA-1
+分子 #5: Nipped-B-like protein
+分子 #8: Transcriptional repressor CTCF
+分子 #6: DNA (118-MER)
+分子 #7: DNA (118-MER)
+分子 #9: ADENOSINE-5'-DIPHOSPHATE
+分子 #10: BERYLLIUM TRIFLUORIDE ION
+分子 #11: ZINC 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: 7.5 |
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グリッド | モデル: Quantifoil R1.2/1.3 / 材質: GOLD / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: HOLEY / 前処理 - タイプ: GLOW DISCHARGE |
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277.15 K |
-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 K3 (6k x 4k) / 撮影したグリッド数: 1 / 平均電子線量: 60.0 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 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
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
初期モデル |
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精密化 | 空間: REAL / プロトコル: RIGID BODY FIT | ||||||||||||
得られたモデル | PDB-7w1m: |