+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-4579 | ||||||||||||
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タイトル | Structure of inner kinetochore CCAN-Cenp-A complex | ||||||||||||
マップデータ | Map of CCAN-Cenp-A | ||||||||||||
試料 |
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キーワード | inner kinetochore / DNA / nucleosome / DNA BINDING PROTEIN | ||||||||||||
機能・相同性 | 機能・相同性情報 2-micrometer circle DNA / 2-micrometer plasmid partitioning / negative regulation of meiotic DNA double-strand break formation involved in reciprocal meiotic recombination / COMA complex / maintenance of meiotic sister chromatid cohesion / PKMTs methylate histone lysines / meiotic sister chromatid segregation / HDMs demethylate histones / Mis6-Sim4 complex / centromere complex assembly ...2-micrometer circle DNA / 2-micrometer plasmid partitioning / negative regulation of meiotic DNA double-strand break formation involved in reciprocal meiotic recombination / COMA complex / maintenance of meiotic sister chromatid cohesion / PKMTs methylate histone lysines / meiotic sister chromatid segregation / HDMs demethylate histones / Mis6-Sim4 complex / centromere complex assembly / HATs acetylate histones / spindle attachment to meiosis I kinetochore / ascospore formation / establishment of meiotic sister chromatid cohesion / Condensation of Prophase Chromosomes / SIRT1 negatively regulates rRNA expression / Activated PKN1 stimulates transcription of AR (androgen receptor) regulated genes KLK2 and KLK3 / attachment of spindle microtubules to kinetochore / Assembly of the ORC complex at the origin of replication / centromeric DNA binding / CENP-A containing chromatin assembly / HDACs deacetylate histones / outer kinetochore / protein localization to chromosome, centromeric region / kinetochore assembly / attachment of mitotic spindle microtubules to kinetochore / Recruitment and ATM-mediated phosphorylation of repair and signaling proteins at DNA double strand breaks / condensed chromosome, centromeric region / establishment of mitotic sister chromatid cohesion / replication fork protection complex / protein localization to kinetochore / RMTs methylate histone arginines / Oxidative Stress Induced Senescence / postreplication repair / spindle pole body / prenyltransferase activity / isoprenoid biosynthetic process / mitotic spindle assembly checkpoint signaling / RNA Polymerase I Promoter Escape / mitotic sister chromatid segregation / Estrogen-dependent gene expression / rRNA transcription / DNA replication initiation / Ub-specific processing proteases / protein localization to CENP-A containing chromatin / CENP-A containing nucleosome / nucleosomal DNA binding / mitotic spindle organization / meiotic cell cycle / chromosome segregation / heterochromatin formation / kinetochore / structural constituent of chromatin / nucleosome / peroxisome / chromatin organization / mitotic cell cycle / sequence-specific DNA binding / protein heterodimerization activity / cell division / DNA repair / protein-containing complex binding / regulation of DNA-templated transcription / structural molecule activity / negative regulation of transcription by RNA polymerase II / DNA binding / nucleus / cytoplasm 類似検索 - 分子機能 | ||||||||||||
生物種 | Saccharomyces cerevisiae (strain ATCC 204508 / S288c) cerevisiae (パン酵母) / Escherichia coli (大腸菌) / Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (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: 4.15 Å | ||||||||||||
データ登録者 | Yan K / Yang J | ||||||||||||
資金援助 | 英国, 3件
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引用 | ジャーナル: Nature / 年: 2019 タイトル: Structure of the inner kinetochore CCAN complex assembled onto a centromeric nucleosome. 著者: Kaige Yan / Jing Yang / Ziguo Zhang / Stephen H McLaughlin / Leifu Chang / Domenico Fasci / Ann E Ehrenhofer-Murray / Albert J R Heck / David Barford / 要旨: In eukaryotes, accurate chromosome segregation in mitosis and meiosis maintains genome stability and prevents aneuploidy. Kinetochores are large protein complexes that, by assembling onto specialized ...In eukaryotes, accurate chromosome segregation in mitosis and meiosis maintains genome stability and prevents aneuploidy. Kinetochores are large protein complexes that, by assembling onto specialized Cenp-A nucleosomes, function to connect centromeric chromatin to microtubules of the mitotic spindle. Whereas the centromeres of vertebrate chromosomes comprise millions of DNA base pairs and attach to multiple microtubules, the simple point centromeres of budding yeast are connected to individual microtubules. All 16 budding yeast chromosomes assemble complete kinetochores using a single Cenp-A nucleosome (Cenp-A), each of which is perfectly centred on its cognate centromere. The inner and outer kinetochore modules are responsible for interacting with centromeric chromatin and microtubules, respectively. Here we describe the cryo-electron microscopy structure of the Saccharomyces cerevisiae inner kinetochore module, the constitutive centromere associated network (CCAN) complex, assembled onto a Cenp-A nucleosome (CCAN-Cenp-A). The structure explains the interdependency of the constituent subcomplexes of CCAN and shows how the Y-shaped opening of CCAN accommodates Cenp-A to enable specific CCAN subunits to contact the nucleosomal DNA and histone subunits. Interactions with the unwrapped DNA duplex at the two termini of Cenp-A are mediated predominantly by a DNA-binding groove in the Cenp-L-Cenp-N subcomplex. Disruption of these interactions impairs assembly of CCAN onto Cenp-A. Our data indicate a mechanism of Cenp-A nucleosome recognition by CCAN and how CCAN acts as a platform for assembly of the outer kinetochore to link centromeres to the mitotic spindle for chromosome segregation. | ||||||||||||
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_4579.map.gz | 10.1 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-4579-v30.xml emd-4579.xml | 38.9 KB 38.9 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_4579.png | 38.6 KB | ||
Filedesc metadata | emd-4579.cif.gz | 8.7 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-4579 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-4579 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_4579_validation.pdf.gz | 219.9 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_4579_full_validation.pdf.gz | 219 KB | 表示 | |
XML形式データ | emd_4579_validation.xml.gz | 6.4 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4579 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4579 | 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_4579.map.gz / 形式: CCP4 / 大きさ: 125 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Map of CCAN-Cenp-A | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.09 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 : Inner kinetochore CCAN complex assembled onto a centromeric nucleosome
+超分子 #1: Inner kinetochore CCAN complex assembled onto a centromeric nucleosome
+超分子 #2: kinetochore
+超分子 #3: DNA
+超分子 #4: Histones
+分子 #1: Inner kinetochore subunit MIF2
+分子 #3: Inner kinetochore subunit MCM16
+分子 #4: Inner kinetochore subunit CTF3
+分子 #6: Inner kinetochore subunit MCM22
+分子 #7: Inner kinetochore subunit IML3
+分子 #8: Inner kinetochore subunit CHL4
+分子 #9: Inner kinetochore subunit MCM21
+分子 #10: Inner kinetochore subunit CTF19
+分子 #11: Inner kinetochore subunit OKP1
+分子 #12: Inner kinetochore subunit AME1
+分子 #13: Inner kinetochore subunit NKP1
+分子 #14: Inner kinetochore subunit NKP2
+分子 #15: Histone H3-like centromeric protein CSE4
+分子 #16: Histone H4
+分子 #17: Histone H2B.2
+分子 #18: Histone H3-like centromeric protein CSE4
+分子 #19: Histone H2A.1
+分子 #20: Histone H2B.1
+分子 #21: Histone H2A.1
+分子 #2: DNA (125-MER)
+分子 #5: DNA (125-MER)
-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 | 1.0 mg/mL |
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緩衝液 | pH: 8 |
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: FEI FALCON III (4k x 4k) 検出モード: COUNTING / 平均電子線量: 32.0 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 |
-画像解析
初期モデル | モデルのタイプ: OTHER / 詳細: Reconstruction using SIMPLE_prime |
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最終 再構成 | 解像度のタイプ: BY AUTHOR / 解像度: 4.15 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 145783 |
初期 角度割当 | タイプ: MAXIMUM LIKELIHOOD |
最終 角度割当 | タイプ: MAXIMUM LIKELIHOOD |