+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 6zz6 | |||||||||
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タイトル | Cryo-EM structure of S.cerevisiae cohesin-Scc2-DNA complex | |||||||||
要素 |
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キーワード | CELL CYCLE / SMC / cohesin / DNA | |||||||||
機能・相同性 | 機能・相同性情報 SMC loading complex / Scc2-Scc4 cohesin loading complex / mitotic cohesin loading / 2-micrometer circle DNA / Establishment of Sister Chromatid Cohesion / Resolution of Sister Chromatid Cohesion / tRNA gene clustering / DNA secondary structure binding / meiotic cohesin complex / establishment of meiotic sister chromatid cohesion ...SMC loading complex / Scc2-Scc4 cohesin loading complex / mitotic cohesin loading / 2-micrometer circle DNA / Establishment of Sister Chromatid Cohesion / Resolution of Sister Chromatid Cohesion / tRNA gene clustering / DNA secondary structure binding / meiotic cohesin complex / establishment of meiotic sister chromatid cohesion / cohesin complex / mitotic cohesin complex / transcription-dependent tethering of RNA polymerase II gene DNA at nuclear periphery / rDNA chromatin condensation / establishment of protein localization to chromatin / synaptonemal complex assembly / SUMOylation of DNA damage response and repair proteins / meiotic sister chromatid cohesion / replication-born double-strand break repair via sister chromatid exchange / establishment of mitotic sister chromatid cohesion / mitotic chromosome condensation / reciprocal meiotic recombination / sister chromatid cohesion / mitotic sister chromatid cohesion / protein acetylation / minor groove of adenine-thymine-rich DNA binding / mitotic sister chromatid segregation / chromosome, centromeric region / protein localization to chromatin / condensed nuclear chromosome / G2/M transition of mitotic cell cycle / double-strand break repair / double-stranded DNA binding / regulation of gene expression / sequence-specific DNA binding / cell division / DNA damage response / chromatin binding / chromatin / apoptotic process / protein kinase binding / ATP hydrolysis activity / mitochondrion / DNA binding / ATP binding / identical protein binding / nucleus / cytosol 類似検索 - 分子機能 | |||||||||
生物種 | Saccharomyces cerevisiae (パン酵母) 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: 3.4 Å | |||||||||
データ登録者 | Lee, B.-G. / Gonzalez Llamazares, A. / Collier, J. / Nasmyth, K.A. / Lowe, J. | |||||||||
資金援助 | 英国, 2件
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引用 | ジャーナル: Elife / 年: 2020 タイトル: Transport of DNA within cohesin involves clamping on top of engaged heads by Scc2 and entrapment within the ring by Scc3. 著者: James E Collier / Byung-Gil Lee / Maurici Brunet Roig / Stanislav Yatskevich / Naomi J Petela / Jean Metson / Menelaos Voulgaris / Andres Gonzalez Llamazares / Jan Löwe / Kim A Nasmyth / 要旨: In addition to extruding DNA loops, cohesin entraps within its SMC-kleisin ring (S-K) individual DNAs during G1 and sister DNAs during S-phase. All three activities require related hook-shaped ...In addition to extruding DNA loops, cohesin entraps within its SMC-kleisin ring (S-K) individual DNAs during G1 and sister DNAs during S-phase. All three activities require related hook-shaped proteins called Scc2 and Scc3. Using thiol-specific crosslinking we provide rigorous proof of entrapment activity in vitro. Scc2 alone promotes entrapment of DNAs in the E-S and E-K compartments, between ATP-bound engaged heads and the SMC hinge and associated kleisin, respectively. This does not require ATP hydrolysis nor is it accompanied by entrapment within S-K rings, which is a slower process requiring Scc3. Cryo-EM reveals that DNAs transported into E-S/E-K compartments are 'clamped' in a sub-compartment created by Scc2's association with engaged heads whose coiled coils are folded around their elbow. We suggest that clamping may be a recurrent feature of cohesin complexes active in loop extrusion and that this conformation precedes the S-K entrapment required for sister chromatid cohesion. | |||||||||
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 | 6zz6.cif.gz | 391.4 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb6zz6.ent.gz | 301.2 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 6zz6.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 | 6zz6_validation.pdf.gz | 1.2 MB | 表示 | wwPDB検証レポート |
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文書・詳細版 | 6zz6_full_validation.pdf.gz | 1.2 MB | 表示 | |
XML形式データ | 6zz6_validation.xml.gz | 61.1 KB | 表示 | |
CIF形式データ | 6zz6_validation.cif.gz | 90.9 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/zz/6zz6 ftp://data.pdbj.org/pub/pdb/validation_reports/zz/6zz6 | 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
-Structural maintenance of chromosomes protein ... , 2種, 2分子 AB
#1: タンパク質 | 分子量: 40854.500 Da / 分子数: 1 / 由来タイプ: 組換発現 詳細: 2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044- ...詳細: 2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224,2-71, 87-195, 1044-1224 由来: (組換発現) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (パン酵母) 株: ATCC 204508 / S288c / 遺伝子: SMC1, CHL10, YFL008W 発現宿主: Spodoptera frugiperda (ツマジロクサヨトウ) 参照: UniProt: P32908 |
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#2: タンパク質 | 分子量: 48295.016 Da / 分子数: 1 / 由来タイプ: 組換発現 詳細: 0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, ...詳細: 0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222,0-224, 997-1071,1104-1222 由来: (組換発現) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (パン酵母) 株: ATCC 204508 / S288c / 遺伝子: SMC3, YJL074C, J1049 発現宿主: Spodoptera frugiperda (ツマジロクサヨトウ) 参照: UniProt: P47037 |
-Sister chromatid cohesion protein ... , 2種, 2分子 CD
#3: タンパク質 | 分子量: 9123.593 Da / 分子数: 1 / 由来タイプ: 組換発現 詳細: 67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, ...詳細: 67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555,67-103, 502-510, 519-555 由来: (組換発現) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (パン酵母) 株: ATCC 204508 / S288c / 遺伝子: MCD1, PDS3, RHC21, SCC1, YDL003W, YD8119.04 発現宿主: Spodoptera frugiperda (ツマジロクサヨトウ) 参照: UniProt: Q12158 |
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#4: タンパク質 | 分子量: 171310.625 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (パン酵母) 株: ATCC 204508 / S288c / 遺伝子: SCC2, YDR180W, YD9395.14 発現宿主: Spodoptera frugiperda (ツマジロクサヨトウ) 参照: UniProt: Q04002 |
-DNA鎖 , 2種, 2分子 FG
#5: DNA鎖 | 分子量: 10604.072 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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#6: DNA鎖 | 分子量: 10297.598 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, 4分子
#7: 化合物 | #8: 化合物 | |
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-詳細
研究の焦点であるリガンドがあるか | N |
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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: 0.485 MDa / 実験値: YES | ||||||||||||||||||||||||
由来(天然) |
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由来(組換発現) |
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緩衝液 | pH: 7.5 | ||||||||||||||||||||||||
試料 | 濃度: 0.3 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | ||||||||||||||||||||||||
試料支持 | グリッドの材料: GOLD / グリッドのサイズ: 200 divisions/in. / グリッドのタイプ: UltrAuFoil | ||||||||||||||||||||||||
急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 100 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 / 倍率(公称値): 81000 X / 最大 デフォーカス(公称値): 3000 nm / 最小 デフォーカス(公称値): 1500 nm / Cs: 2.7 mm / C2レンズ絞り径: 50 µm / 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: 55 e/Å2 / フィルム・検出器のモデル: GATAN K3 (6k x 4k) |
-解析
EMソフトウェア | 名称: EPU / カテゴリ: 画像取得 |
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CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION |
3次元再構成 | 解像度: 3.4 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 588164 / 対称性のタイプ: POINT |