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- EMDB-27663: Open state of RFC:PCNA bound to a 3' ss/dsDNA junction -

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基本情報

登録情報
データベース: EMDB / ID: EMD-27663
タイトルOpen state of RFC:PCNA bound to a 3' ss/dsDNA junction
マップデータOpen RFC-PCNA
試料
  • 複合体: Open state of RFC:PCNA bound to a 3' ss/dsDNA junction
    • タンパク質・ペプチド: x 6種
    • DNA: x 3種
  • リガンド: x 4種
キーワードREPLICATION-DNA complex
機能・相同性
機能・相同性情報


DNA clamp unloader activity / DNA clamp unloading / Mismatch repair (MMR) directed by MSH2:MSH6 (MutSalpha) / Gap-filling DNA repair synthesis and ligation in GG-NER / meiotic mismatch repair / Processive synthesis on the lagging strand / Removal of the Flap Intermediate / Ctf18 RFC-like complex / Rad17 RFC-like complex / DNA replication factor C complex ...DNA clamp unloader activity / DNA clamp unloading / Mismatch repair (MMR) directed by MSH2:MSH6 (MutSalpha) / Gap-filling DNA repair synthesis and ligation in GG-NER / meiotic mismatch repair / Processive synthesis on the lagging strand / Removal of the Flap Intermediate / Ctf18 RFC-like complex / Rad17 RFC-like complex / 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 / DNA clamp loader activity / 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 / replication fork / positive regulation of DNA replication / DNA damage checkpoint signaling / 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
Replication factor C subunit 1 / DNA replication factor RFC1, C-terminal / Replication factor RFC1 C terminal domain / Replication factor C, C-terminal / Replication factor C C-terminal domain / : / DNA polymerase III, delta subunit / DNA polymerase III, clamp loader complex, gamma/delta/delta subunit, C-terminal / Proliferating cell nuclear antigen signature 2. / Proliferating cell nuclear antigen, PCNA, conserved site ...Replication factor C subunit 1 / DNA replication factor RFC1, C-terminal / Replication factor RFC1 C terminal domain / Replication factor C, C-terminal / Replication factor C C-terminal domain / : / DNA polymerase III, delta subunit / DNA polymerase III, clamp loader complex, gamma/delta/delta subunit, C-terminal / Proliferating cell nuclear antigen signature 2. / Proliferating cell nuclear antigen, PCNA, conserved site / Proliferating cell nuclear antigen signature 1. / Proliferating cell nuclear antigen, PCNA / Proliferating cell nuclear antigen, PCNA, N-terminal / Proliferating cell nuclear antigen, PCNA, C-terminal / Proliferating cell nuclear antigen, N-terminal domain / Proliferating cell nuclear antigen, C-terminal domain / : / BRCA1 C Terminus (BRCT) domain / breast cancer carboxy-terminal domain / BRCT domain profile. / BRCT domain / BRCT domain superfamily / ATPase family associated with various cellular activities (AAA) / ATPase, AAA-type, core / ATPases associated with a variety of cellular activities / AAA+ ATPase domain / P-loop containing nucleoside triphosphate hydrolase
類似検索 - ドメイン・相同性
Proliferating cell nuclear antigen / Replication factor C subunit 5 / Replication factor C subunit 3 / Replication factor C subunit 1 / Replication factor C subunit 4 / Replication factor C subunit 2
類似検索 - 構成要素
生物種Saccharomyces cerevisiae (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: 2.1 Å
データ登録者Schrecker M / Hite RK
資金援助 米国, 3件
OrganizationGrant number国
National Institutes of Health/National Cancer Institute (NIH/NCI)CA008748 米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM107239 米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM127428 米国
引用ジャーナル: Elife / 年: 2022
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
登録2022年7月20日-
ヘッダ(付随情報) 公開2022年8月24日-
マップ公開2022年8月24日-
更新2024年2月14日-
現状2024年2月14日処理サイト: RCSB / 状態: 公開

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構造の表示

添付画像

emd_27663.png

ダウンロードとリンク

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マップ

ファイルダウンロード / ファイル: emd_27663.map.gz / 形式: CCP4 / 大きさ: 216 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈Open RFC-PCNA
ボクセルのサイズX=Y=Z: 0.826 Å
密度
表面レベル登録者による: 0.2
最小 - 最大-0.65756357 - 3.127737
平均 (標準偏差)0.000001441125 (±0.05148439)
対称性空間群: 1
詳細

EMDB XML:

マップ形状
Axis orderXYZ
Origin000
サイズ384384384
Spacing384384384
セルA=B=C: 317.184 Å
α=β=γ: 90.0 °

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添付データ

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追加マップ: Density modified and 1.5x resampled

ファイルemd_27663_additional_1.map
注釈Density modified and 1.5x resampled
投影像・断面図
軸ZYX

投影像

断面 (1/2)
密度ヒストグラム

ヒストグラム

ヒストグラム (対数)

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ハーフマップ: A

ファイルemd_27663_half_map_1.map
注釈A
投影像・断面図
軸ZYX

投影像

断面 (1/2)
密度ヒストグラム

ヒストグラム

ヒストグラム (対数)

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ハーフマップ: B

ファイルemd_27663_half_map_2.map
注釈B
投影像・断面図
軸ZYX

投影像

断面 (1/2)
密度ヒストグラム

ヒストグラム

ヒストグラム (対数)

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試料の構成要素

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全体 : Open state of RFC:PCNA bound to a 3' ss/dsDNA junction

全体名称: Open state of RFC:PCNA bound to a 3' ss/dsDNA junction
要素
  • 複合体: Open state of RFC:PCNA bound to a 3' ss/dsDNA junction
    • タンパク質・ペプチド: Replication factor C subunit 1
    • タンパク質・ペプチド: Replication factor C subunit 4
    • タンパク質・ペプチド: Replication factor C subunit 3
    • タンパク質・ペプチド: Replication factor C subunit 2
    • タンパク質・ペプチド: Replication factor C subunit 5
    • タンパク質・ペプチド: Proliferating cell nuclear antigen
    • DNA: DNA (5'-D(*TP*TP*TP*TP*TP*T)-3')
    • DNA: DNA (5'-D(P*TP*CP*CP*GP*AP*GP*CP*GP*AP*A)-3')
    • DNA: DNA (5'-D(P*TP*TP*TP*GP*CP*CP*CP*GP*GP*A)-3')
  • リガンド: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER
  • リガンド: MAGNESIUM ION
  • リガンド: GUANOSINE-5'-DIPHOSPHATE
  • リガンド: water

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超分子 #1: Open state of RFC:PCNA bound to a 3' ss/dsDNA junction

超分子名称: Open state of RFC:PCNA bound to a 3' ss/dsDNA junction
タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#9
由来(天然)生物種: Saccharomyces cerevisiae (パン酵母)

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分子 #1: Replication factor C subunit 1

分子名称: Replication factor C subunit 1 / タイプ: protein_or_peptide / ID: 1 / コピー数: 1 / 光学異性体: LEVO
由来(天然)生物種: Saccharomyces cerevisiae (パン酵母)
分子量理論値: 95.048195 KDa
組換発現生物種: Saccharomyces cerevisiae (パン酵母)
配列文字列: MVNISDFFGK NKKSVRSSTS RPTRQVGSSK PEVIDLDTES DQESTNKTPK KMPVSNVIDV SETPEGEKKL PLPAKRKASS PTVKPASSK KTKPSSKSSD SASNITAQDV LDKIPSLDLS NVHVKENAKF DFKSANSNAD PDEIVSEIGS FPEGKPNCLL G LTIVFTGV ...文字列:
MVNISDFFGK NKKSVRSSTS RPTRQVGSSK PEVIDLDTES DQESTNKTPK KMPVSNVIDV SETPEGEKKL PLPAKRKASS PTVKPASSK KTKPSSKSSD SASNITAQDV LDKIPSLDLS NVHVKENAKF DFKSANSNAD PDEIVSEIGS FPEGKPNCLL G LTIVFTGV LPTLERGASE ALAKRYGARV TKSISSKTSV VVLGDEAGPK KLEKIKQLKI KAIDEEGFKQ LIAGMPAEGG DG EAAEKAR RKLEEQHNIA TKEAELLVKK EEERSKKLAA TRVSGGHLER DNVVREEDKL WTVKYAPTNL QQVCGNKGSV MKL KNWLAN WENSKKNSFK HAGKDGSGVF RAAMLYGPPG IGKTTAAHLV AQELGYDILE QNASDVRSKT LLNAGVKNAL DNMS VVGYF KHNEEAQNLN GKHFVIIMDE VDGMSGGDRG GVGQLAQFCR KTSTPLILIC NERNLPKMRP FDRVCLDIQF RRPDA NSIK SRLMTIAIRE KFKLDPNVID RLIQTTRGDI RQVINLLSTI STTTKTINHE NINEISKAWE KNIALKPFDI AHKMLD GQI YSDIGSRNFT LNDKIALYFD DFDFTPLMIQ ENYLSTRPSV LKPGQSHLEA VAEAANCISL GDIVEKKIRS SEQLWSL LP LHAVLSSVYP ASKVAGHMAG RINFTAWLGQ NSKSAKYYRL LQEIHYHTRL GTSTDKIGLR LDYLPTFRKR LLDPFLKQ G ADAISSVIEV MDDYYLTKED WDSIMEFFVG PDVTTAIIKK IPATVKSGFT RKYNSMTHPV AIYRTGSTIG GGGVGTSTS TPDFEDVVDA DDNPVPADDE ETQDSSTDLK KDKLIKQKAK PTKRKTATSK PGGSKKRKTK A

UniProtKB: Replication factor C subunit 1

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分子 #2: Replication factor C subunit 4

分子名称: Replication factor C subunit 4 / タイプ: protein_or_peptide / ID: 2 / コピー数: 1 / 光学異性体: LEVO
由来(天然)生物種: Saccharomyces cerevisiae (パン酵母)
分子量理論値: 36.201039 KDa
組換発現生物種: