+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 6fml | ||||||||||||
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タイトル | CryoEM Structure INO80core Nucleosome complex | ||||||||||||
要素 |
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キーワード | DNA BINDING PROTEIN / INO80 / Nucleosome / ATP dependent Chromatin Remodeller | ||||||||||||
機能・相同性 | 機能・相同性情報 HDACs deacetylate histones / HATs acetylate histones / RMTs methylate histone arginines / DASH complex / protein transport along microtubule to mitotic spindle pole body / mitotic sister chromatid biorientation / Metalloprotease DUBs / UCH proteinases / Ub-specific processing proteases / attachment of spindle microtubules to kinetochore ...HDACs deacetylate histones / HATs acetylate histones / RMTs methylate histone arginines / DASH complex / protein transport along microtubule to mitotic spindle pole body / mitotic sister chromatid biorientation / Metalloprotease DUBs / UCH proteinases / Ub-specific processing proteases / attachment of spindle microtubules to kinetochore / attachment of mitotic spindle microtubules to kinetochore / Ino80 complex / negative regulation of megakaryocyte differentiation / ATP-dependent activity, acting on DNA / protein localization to CENP-A containing chromatin / Chromatin modifying enzymes / Replacement of protamines by nucleosomes in the male pronucleus / CENP-A containing nucleosome / Packaging Of Telomere Ends / Recognition and association of DNA glycosylase with site containing an affected purine / Cleavage of the damaged purine / Deposition of new CENPA-containing nucleosomes at the centromere / nucleosomal DNA binding / Recognition and association of DNA glycosylase with site containing an affected pyrimidine / Cleavage of the damaged pyrimidine / Inhibition of DNA recombination at telomere / telomere organization / Meiotic synapsis / Interleukin-7 signaling / RNA Polymerase I Promoter Opening / Assembly of the ORC complex at the origin of replication / SUMOylation of chromatin organization proteins / Regulation of endogenous retroelements by the Human Silencing Hub (HUSH) complex / DNA methylation / Condensation of Prophase Chromosomes / SIRT1 negatively regulates rRNA expression / Chromatin modifications during the maternal to zygotic transition (MZT) / ERCC6 (CSB) and EHMT2 (G9a) positively regulate rRNA expression / HCMV Late Events / innate immune response in mucosa / PRC2 methylates histones and DNA / Regulation of endogenous retroelements by KRAB-ZFP proteins / Defective pyroptosis / Regulation of endogenous retroelements by Piwi-interacting RNAs (piRNAs) / HDACs deacetylate histones / Nonhomologous End-Joining (NHEJ) / helicase activity / RNA Polymerase I Promoter Escape / Transcriptional regulation by small RNAs / Formation of the beta-catenin:TCF transactivating complex / RUNX1 regulates genes involved in megakaryocyte differentiation and platelet function / NoRC negatively regulates rRNA expression / Activated PKN1 stimulates transcription of AR (androgen receptor) regulated genes KLK2 and KLK3 / G2/M DNA damage checkpoint / HDMs demethylate histones / B-WICH complex positively regulates rRNA expression / DNA Damage/Telomere Stress Induced Senescence / heterochromatin formation / mitotic spindle / PKMTs methylate histone lysines / Metalloprotease DUBs / Meiotic recombination / kinetochore / Pre-NOTCH Transcription and Translation / RMTs methylate histone arginines / Activation of anterior HOX genes in hindbrain development during early embryogenesis / HCMV Early Events / Transcriptional regulation of granulopoiesis / structural constituent of chromatin / UCH proteinases / antimicrobial humoral immune response mediated by antimicrobial peptide / nucleosome / nucleosome assembly / E3 ubiquitin ligases ubiquitinate target proteins / antibacterial humoral response / Recruitment and ATM-mediated phosphorylation of repair and signaling proteins at DNA double strand breaks / chromatin organization / RUNX1 regulates transcription of genes involved in differentiation of HSCs / Factors involved in megakaryocyte development and platelet production / HATs acetylate histones / Processing of DNA double-strand break ends / Senescence-Associated Secretory Phenotype (SASP) / DNA helicase / Oxidative Stress Induced Senescence / Estrogen-dependent gene expression / chromosome, telomeric region / Ub-specific processing proteases / defense response to Gram-positive bacterium / chromatin remodeling / protein heterodimerization activity / Amyloid fiber formation / DNA repair / enzyme binding / ATP hydrolysis activity / protein-containing complex / DNA binding / RNA binding / extracellular space / extracellular exosome / extracellular region 類似検索 - 分子機能 | ||||||||||||
生物種 | Chaetomium thermophilum (菌類) Chaetomium thermophilum var. thermophilum DSM 1495 (菌類) Homo sapiens (ヒト) synthetic construct (人工物) | ||||||||||||
手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 4.34 Å | ||||||||||||
データ登録者 | Eustermann, S. / Schall, K. / Kostrewa, D. / Strauss, M. / Hopfner, K. | ||||||||||||
資金援助 | ドイツ, 3件
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引用 | ジャーナル: Nature / 年: 2018 タイトル: Structural basis for ATP-dependent chromatin remodelling by the INO80 complex. 著者: Sebastian Eustermann / Kevin Schall / Dirk Kostrewa / Kristina Lakomek / Mike Strauss / Manuela Moldt / Karl-Peter Hopfner / 要旨: In the eukaryotic nucleus, DNA is packaged in the form of nucleosomes, each of which comprises about 147 base pairs of DNA wrapped around a histone protein octamer. The position and histone ...In the eukaryotic nucleus, DNA is packaged in the form of nucleosomes, each of which comprises about 147 base pairs of DNA wrapped around a histone protein octamer. The position and histone composition of nucleosomes is governed by ATP-dependent chromatin remodellers such as the 15-subunit INO80 complex . INO80 regulates gene expression, DNA repair and replication by sliding nucleosomes, the exchange of histone H2A.Z with H2A, and the positioning of + 1 and -1 nucleosomes at promoter DNA. The structures and mechanisms of these remodelling reactions are currently unknown. Here we report the cryo-electron microscopy structure of the evolutionarily conserved core of the INO80 complex from the fungus Chaetomium thermophilum bound to a nucleosome, at a global resolution of 4.3 Å and with major parts at 3.7 Å. The INO80 core cradles one entire gyre of the nucleosome through multivalent DNA and histone contacts. An Rvb1/Rvb2 AAA ATPase heterohexamer is an assembly scaffold for the complex and acts as a 'stator' for the motor and nucleosome-gripping subunits. The Swi2/Snf2 ATPase motor binds to nucleosomal DNA at superhelical location -6, unwraps approximately 15 base pairs, disrupts the H2A-DNA contacts and is poised to pump entry DNA into the nucleosome. Arp5 and Ies6 bind superhelical locations -2 and -3 to act as a counter grip for the motor, on the other side of the H2A-H2B dimer. The Arp5 insertion domain forms a grappler element that binds the nucleosome dyad, connects the Arp5 actin-fold and entry DNA over a distance of about 90 Å and packs against histone H2A-H2B near the 'acidic patch'. Our structure together with biochemical data suggests a unified mechanism for nucleosome sliding and histone editing by INO80. The motor is part of a macromolecular ratchet, persistently pumping entry DNA across the H2A-H2B dimer against the Arp5 grip until a large nucleosome translocation step occurs. The transient exposure of H2A-H2B by motor activity as well as differential recognition of H2A.Z and H2A may regulate histone exchange. | ||||||||||||
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 | 分子: MolmilJmol/JSmol |
-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 6fml.cif.gz | 1006.8 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb6fml.ent.gz | 789.7 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 6fml.json.gz | ツリー表示 | PDBx/mmJSON形式 | |
その他 | その他のダウンロード |
-検証レポート
文書・要旨 | 6fml_validation.pdf.gz | 1.4 MB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 6fml_full_validation.pdf.gz | 1.4 MB | 表示 | |
XML形式データ | 6fml_validation.xml.gz | 131.6 KB | 表示 | |
CIF形式データ | 6fml_validation.cif.gz | 204.7 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/fm/6fml ftp://data.pdbj.org/pub/pdb/validation_reports/fm/6fml | HTTPS FTP |
-関連構造データ
-リンク
-集合体
登録構造単位 |
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-要素
-タンパク質 , 10種, 18分子 ABCDEFGHIJMQNROSPT
#1: タンパク質 | 分子量: 50451.848 Da / 分子数: 3 / 由来タイプ: 組換発現 由来: (組換発現) Chaetomium thermophilum (strain DSM 1495 / CBS 144.50 / IMI 039719) (菌類) 遺伝子: CTHT_0006820 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: G0RYI5, DNA helicase #2: タンパク質 | 分子量: 53212.746 Da / 分子数: 3 / 由来タイプ: 組換発現 由来: (組換発現) Chaetomium thermophilum (strain DSM 1495 / CBS 144.50 / IMI 039719) (菌類) 遺伝子: CTHT_0006170 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: G0RYC2, DNA helicase #3: タンパク質 | | 分子量: 210443.344 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) Chaetomium thermophilum (strain DSM 1495 / CBS 144.50 / IMI 039719) (菌類) 発現宿主: Trichoplusia ni (イラクサキンウワバ) #4: タンパク質 | | 分子量: 52018.512 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) Chaetomium thermophilum (strain DSM 1495 / CBS 144.50 / IMI 039719) (菌類), (組換発現) Chaetomium thermophilum var. thermophilum DSM 1495 (菌類) 遺伝子: CTHT_0004910 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: G0RY01 #5: タンパク質 | | 分子量: 23127.523 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) Chaetomium thermophilum (strain DSM 1495 / CBS 144.50 / IMI 039719) (菌類) 遺伝子: CTHT_0032670 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: G0S590 #6: タンパク質 | | 分子量: 85996.453 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) Chaetomium thermophilum (strain DSM 1495 / CBS 144.50 / IMI 039719) (菌類), (組換発現) Chaetomium thermophilum var. thermophilum DSM 1495 (菌類) 遺伝子: CTHT_0032660 / 発現宿主: Trichoplusia ni (イラクサキンウワバ) / 参照: UniProt: G0S589 #9: タンパク質 | 分子量: 15289.904 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: HIST2H3A, HIST2H3C, H3F2, H3FM, HIST2H3D / 発現宿主: Escherichia coli (大腸菌) / 参照: UniProt: Q71DI3 #10: タンパク質 | 分子量: 11263.231 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) 遺伝子: HIST1H4A, H4/A, H4FA, HIST1H4B, H4/I, H4FI, HIST1H4C, H4/G, H4FG, HIST1H4D, H4/B, H4FB, HIST1H4E, H4/J, H4FJ, HIST1H4F, H4/C, H4FC, HIST1H4H, H4/H, H4FH, HIST1H4I, H4/M, H4FM, HIST1H4J, ...遺伝子: HIST1H4A, H4/A, H4FA, HIST1H4B, H4/I, H4FI, HIST1H4C, H4/G, H4FG, HIST1H4D, H4/B, H4FB, HIST1H4E, H4/J, H4FJ, HIST1H4F, H4/C, H4FC, HIST1H4H, H4/H, H4FH, HIST1H4I, H4/M, H4FM, HIST1H4J, H4/E, H4FE, HIST1H4K, H4/D, H4FD, HIST1H4L, H4/K, H4FK, HIST2H4A, H4/N, H4F2, H4FN, HIST2H4, HIST2H4B, H4/O, H4FO, HIST4H4 発現宿主: Escherichia coli (大腸菌) / 参照: UniProt: P62805 #11: タンパク質 | 分子量: 13990.342 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) / 発現宿主: Escherichia coli (大腸菌) / 参照: UniProt: P0C0S9, UniProt: P0C0S8*PLUS #12: タンパク質 | 分子量: 13806.018 Da / 分子数: 2 / 由来タイプ: 組換発現 / 由来: (組換発現) Homo sapiens (ヒト) 遺伝子: HIST1H2BC, H2BFL, HIST1H2BE, H2BFH, HIST1H2BF, H2BFG, HIST1H2BG, H2BFA, HIST1H2BI, H2BFK 発現宿主: Escherichia coli (大腸菌) / 参照: UniProt: P62807 |
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-Nucleosomal DNA Strand ... , 2種, 2分子 KL
#7: DNA鎖 | 分子量: 60652.645 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) synthetic construct (人工物) |
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#8: DNA鎖 | 分子量: 60376.426 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) synthetic construct (人工物) |
-非ポリマー , 2種, 7分子
#13: 化合物 | ChemComp-ADP / #14: 化合物 | ChemComp-ATP / | |
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-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
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EM実験 | 試料の集合状態: PARTICLE / 3次元再構成法: 単粒子再構成法 |
-試料調製
構成要素 |
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分子量 | 値: 1 MDa / 実験値: NO | ||||||||||||||||||||||||||||||
由来(天然) |
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由来(組換発現) |
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緩衝液 | pH: 8 詳細: 20 mM HEPES pH 8, 60 mM KCl, 0.5% glycerol, 0.25 mM CaCl2, 20 uM ZnCl2, 0.25 mM DTT, 0.05% Octyl-beta-glucoside | ||||||||||||||||||||||||||||||
試料 | 濃度: 1 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES 詳細: Monodisperse sample: INO80core complex reconstituted with nucleosomal substrate was purified by gelfiltration. Addition of nucleotides or crosslinking was not required. | ||||||||||||||||||||||||||||||
試料支持 | グリッドの材料: COPPER / グリッドのサイズ: 200 divisions/in. / グリッドのタイプ: Quantifoil R2/1 | ||||||||||||||||||||||||||||||
急速凍結 | 装置: LEICA EM GP / 凍結剤: ETHANE / 湿度: 95 % / 凍結前の試料温度: 281 K |
-電子顕微鏡撮影
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TITAN KRIOS |
電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: FLOOD BEAM |
電子レンズ | モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 3500 nm / Calibrated defocus min: 1300 nm / アライメント法: COMA FREE |
試料ホルダ | 凍結剤: NITROGEN 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
撮影 | 電子線照射量: 59.6 e/Å2 / 検出モード: COUNTING フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 撮影したグリッド数: 1 / 実像数: 3992 詳細: Images were collected in movie mode with 4 frames per second and 10s total aquisition |
画像スキャン | 動画フレーム数/画像: 40 / 利用したフレーム数/画像: 1-40 |
-解析
ソフトウェア | 名称: PHENIX / バージョン: 1.13_2998: / 分類: 精密化 | ||||||||||||||||||||||||||||||||||||
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EMソフトウェア |
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CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||
粒子像の選択 | 選択した粒子像数: 251692 | ||||||||||||||||||||||||||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||||||||||||||||||||||||||
3次元再構成 | 解像度: 4.34 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 33937 / 対称性のタイプ: POINT |