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データを開く
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基本情報
登録情報 | ![]() | ||||||||||||
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タイトル | Intermediate state of RFC:PCNA bound to a 3' ss/dsDNA junction | ||||||||||||
![]() | RFC-PCNA intermediate state | ||||||||||||
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![]() | REPLICATION-DNA complex | ||||||||||||
機能・相同性 | ![]() DNA clamp unloading / Mismatch repair (MMR) directed by MSH2:MSH6 (MutSalpha) / Gap-filling DNA repair synthesis and ligation in GG-NER / meiotic mismatch repair / Rad17 RFC-like complex / Processive synthesis on the lagging strand / Removal of the Flap Intermediate / Ctf18 RFC-like complex / DNA clamp loader activity / DNA replication factor C complex ...DNA clamp unloading / Mismatch repair (MMR) directed by MSH2:MSH6 (MutSalpha) / Gap-filling DNA repair synthesis and ligation in GG-NER / meiotic mismatch repair / Rad17 RFC-like complex / Processive synthesis on the lagging strand / Removal of the Flap Intermediate / Ctf18 RFC-like complex / DNA clamp loader activity / DNA replication factor C complex / Elg1 RFC-like complex / E3 ubiquitin ligases ubiquitinate target proteins / Polymerase switching / SUMOylation of DNA replication proteins / positive regulation of DNA metabolic process / maintenance of DNA trinucleotide repeats / Translesion Synthesis by POLH / Translesion synthesis by REV1 / Translesion synthesis by POLK / Translesion synthesis by POLI / establishment of mitotic sister chromatid cohesion / DNA replication checkpoint signaling / PCNA complex / Activation of ATR in response to replication stress / Termination of translesion DNA synthesis / lagging strand elongation / postreplication repair / sister chromatid cohesion / silent mating-type cassette heterochromatin formation / mitotic sister chromatid cohesion / error-free translesion synthesis / DNA polymerase processivity factor activity / leading strand elongation / Gap-filling DNA repair synthesis and ligation in TC-NER / Dual incision in TC-NER / subtelomeric heterochromatin formation / mismatch repair / translesion synthesis / positive regulation of DNA repair / positive regulation of DNA replication / DNA damage checkpoint signaling / replication fork / nucleotide-excision repair / DNA-templated DNA replication / mitotic cell cycle / chromosome, telomeric region / cell division / DNA repair / ATP hydrolysis activity / DNA binding / ATP binding / identical protein binding / nucleus / cytosol 類似検索 - 分子機能 | ||||||||||||
生物種 | ![]() ![]() | ||||||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 2.92 Å | ||||||||||||
![]() | Schrecker M / Hite RK | ||||||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Multistep loading of a DNA sliding clamp onto DNA by replication factor C. 著者: Marina Schrecker / Juan C Castaneda / Sujan Devbhandari / Charanya Kumar / Dirk Remus / Richard K Hite / ![]() 要旨: The DNA sliding clamp proliferating cell nuclear antigen (PCNA) is an essential co-factor for many eukaryotic DNA metabolic enzymes. PCNA is loaded around DNA by the ATP-dependent clamp loader ...The DNA sliding clamp proliferating cell nuclear antigen (PCNA) is an essential co-factor for many eukaryotic DNA metabolic enzymes. PCNA is loaded around DNA by the ATP-dependent clamp loader replication factor C (RFC), which acts at single-stranded (ss)/double-stranded DNA (dsDNA) junctions harboring a recessed 3' end (3' ss/dsDNA junctions) and at DNA nicks. To illuminate the loading mechanism we have investigated the structure of RFC:PCNA bound to ATPγS and 3' ss/dsDNA junctions or nicked DNA using cryogenic electron microscopy. Unexpectedly, we observe open and closed PCNA conformations in the RFC:PCNA:DNA complex, revealing that PCNA can adopt an open, planar conformation that allows direct insertion of dsDNA, and raising the question of whether PCNA ring closure is mechanistically coupled to ATP hydrolysis. By resolving multiple DNA-bound states of RFC:PCNA we observe that partial melting facilitates lateral insertion into the central channel formed by RFC:PCNA. We also resolve the Rfc1 N-terminal domain and demonstrate that its single BRCT domain participates in coordinating DNA prior to insertion into the central RFC channel, which promotes PCNA loading on the lagging strand of replication forks in vitro. Combined, our data suggest a comprehensive and fundamentally revised model for the RFC-catalyzed loading of PCNA onto DNA. | ||||||||||||
履歴 |
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構造の表示
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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 | ![]() | 108.2 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 29.8 KB 29.8 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 12.7 KB | 表示 | ![]() |
画像 | ![]() | 183.1 KB | ||
Filedesc metadata | ![]() | 8.1 KB | ||
その他 | ![]() ![]() ![]() | 676.6 MB 200.3 MB 200.3 MB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
関連構造データ | ![]() 8dqzMC ![]() 8dqwC ![]() 8dqxC ![]() 8dr0C ![]() 8dr1C ![]() 8dr3C ![]() 8dr4C ![]() 8dr5C ![]() 8dr6C ![]() 8dr7C C: 同じ文献を引用 ( M: このマップから作成された原子モデル |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
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リンク
EMDBのページ | ![]() ![]() |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||
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注釈 | RFC-PCNA intermediate state | ||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 0.826 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
-追加マップ: Density modified and 1.5x sampled map
ファイル | emd_27666_additional_1.map | ||||||||||||
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注釈 | Density modified and 1.5x sampled map | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: A
ファイル | emd_27666_half_map_1.map | ||||||||||||
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注釈 | A | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: B
ファイル | emd_27666_half_map_2.map | ||||||||||||
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注釈 | B | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
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試料の構成要素
+全体 : Intermediate state of RFC:PCNA bound to a 3' ss/dsDNA junction
+超分子 #1: Intermediate state of RFC:PCNA bound to a 3' ss/dsDNA junction
+分子 #1: Replication factor C subunit 1
+分子 #2: Replication factor C subunit 4
+分子 #3: Replication factor C subunit 3
+分子 #4: Replication factor C subunit 2
+分子 #5: Replication factor C subunit 5
+分子 #6: Proliferating cell nuclear antigen
+分子 #7: DNA (5'-D(P*TP*TP*TP*TP*TP*TP*CP*GP*GP*GP*GP*GP*GP*GP*CP*CP*CP*CP...
+分子 #8: DNA (5'-D(P*CP*CP*CP*CP*GP*GP*GP*GP*CP*CP*CP*CP*CP*CP*CP*GP*GP*C)-3')
+分子 #9: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER
+分子 #10: MAGNESIUM ION
+分子 #11: GUANOSINE-5'-DIPHOSPHATE
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
濃度 | 1.0 mg/mL | ||||||||
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緩衝液 | pH: 7.6 構成要素:
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グリッド | モデル: Quantifoil / 材質: GRAPHENE OXIDE / メッシュ: 400 / 支持フィルム - 材質: GRAPHENE OXIDE / 支持フィルム - トポロジー: CONTINUOUS | ||||||||
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 297 K / 装置: FEI VITROBOT MARK IV |
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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) / 平均電子線量: 66.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 2.0 µm 最小 デフォーカス(公称値): 0.7000000000000001 µm |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |