+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-22803 | ||||||||||||
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タイトル | Cryo-EM 3D map of the Saccharomyces cerevisiae replicative polymerase delta in complex with a primer/template and the PCNA clamp | ||||||||||||
マップデータ | Replicative polymerase delta in complex with a primer/template and the PCNA clamp | ||||||||||||
試料 |
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機能・相同性 | 機能・相同性情報 delta DNA polymerase complex / H3-H4 histone complex chaperone activity / DNA amplification / DNA-templated DNA replication maintenance of fidelity / zeta DNA polymerase complex / RNA-templated DNA biosynthetic process / Mismatch repair (MMR) directed by MSH2:MSH6 (MutSalpha) / meiotic mismatch repair / Processive synthesis on the lagging strand / Removal of the Flap Intermediate ...delta DNA polymerase complex / H3-H4 histone complex chaperone activity / DNA amplification / DNA-templated DNA replication maintenance of fidelity / zeta DNA polymerase complex / RNA-templated DNA biosynthetic process / Mismatch repair (MMR) directed by MSH2:MSH6 (MutSalpha) / meiotic mismatch repair / Processive synthesis on the lagging strand / Removal of the Flap Intermediate / E3 ubiquitin ligases ubiquitinate target proteins / Polymerase switching / DNA replication, removal of RNA primer / positive regulation of DNA metabolic process / SUMOylation of DNA replication proteins / maintenance of DNA trinucleotide repeats / nucleotide-excision repair, DNA gap filling / 3'-5'-DNA exonuclease activity / Translesion Synthesis by POLH / Translesion synthesis by REV1 / Translesion synthesis by POLK / Translesion synthesis by POLI / PCNA complex / DNA replication proofreading / establishment of mitotic sister chromatid cohesion / Termination of translesion DNA synthesis / lagging strand elongation / double-strand break repair via break-induced replication / postreplication repair / DNA strand elongation involved in DNA replication / silent mating-type cassette heterochromatin formation / 加水分解酵素; エステル加水分解酵素; 5'-リン酸モノエステル産生エンドデオキシリボヌクレアーゼ / DNA metabolic process / mitotic sister chromatid cohesion / error-free translesion synthesis / leading strand elongation / DNA polymerase processivity factor activity / regulation of DNA replication / Dual incision in TC-NER / subtelomeric heterochromatin formation / error-prone translesion synthesis / mismatch repair / translesion synthesis / base-excision repair, gap-filling / positive regulation of DNA repair / positive regulation of DNA replication / replication fork / nucleotide-excision repair / base-excision repair / DNA-templated DNA replication / double-strand break repair via nonhomologous end joining / mitotic cell cycle / 4 iron, 4 sulfur cluster binding / DNA replication / DNA-directed DNA polymerase / molecular adaptor activity / chromosome, telomeric region / DNA-directed DNA polymerase activity / nucleotide binding / DNA binding / identical protein binding / nucleus / metal ion binding / cytosol 類似検索 - 分子機能 | ||||||||||||
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: 3.2 Å | ||||||||||||
データ登録者 | Zheng F / Georgescu R / Li H / O'Donnell ME | ||||||||||||
資金援助 | 米国, 3件
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引用 | ジャーナル: Proc Natl Acad Sci U S A / 年: 2020 タイトル: Structure of eukaryotic DNA polymerase δ bound to the PCNA clamp while encircling DNA. 著者: Fengwei Zheng / Roxana E Georgescu / Huilin Li / Michael E O'Donnell / 要旨: The DNA polymerase (Pol) δ of (S.c.) is composed of the catalytic subunit Pol3 along with two regulatory subunits, Pol31 and Pol32. Pol δ binds to proliferating cell nuclear antigen (PCNA) and ...The DNA polymerase (Pol) δ of (S.c.) is composed of the catalytic subunit Pol3 along with two regulatory subunits, Pol31 and Pol32. Pol δ binds to proliferating cell nuclear antigen (PCNA) and functions in genome replication, repair, and recombination. Unique among DNA polymerases, the Pol3 catalytic subunit contains a 4Fe-4S cluster that may sense the cellular redox state. Here we report the 3.2-Å cryo-EM structure of S.c. Pol δ in complex with primed DNA, an incoming ddTTP, and the PCNA clamp. Unexpectedly, Pol δ binds only one subunit of the PCNA trimer. This singular yet extensive interaction holds DNA such that the 2-nm-wide DNA threads through the center of the 3-nm interior channel of the clamp without directly contacting the protein. Thus, a water-mediated clamp and DNA interface enables the PCNA clamp to "waterskate" along the duplex with minimum drag. Pol31 and Pol32 are positioned off to the side of the catalytic Pol3-PCNA-DNA axis. We show here that Pol31-Pol32 binds single-stranded DNA that we propose underlies polymerase recycling during lagging strand synthesis, in analogy to replicase. Interestingly, the 4Fe-4S cluster in the C-terminal CysB domain of Pol3 forms the central interface to Pol31-Pol32, and this strategic location may explain the regulation of the oxidation state on Pol δ activity, possibly useful during cellular oxidative stress. Importantly, human cancer and other disease mutations map to nearly every domain of Pol3, suggesting that all aspects of Pol δ replication are important to human health and disease. | ||||||||||||
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_22803.map.gz | 267.1 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-22803-v30.xml emd-22803.xml | 19.1 KB 19.1 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_22803.png | 157.6 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-22803 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-22803 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_22803_validation.pdf.gz | 453.8 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_22803_full_validation.pdf.gz | 453.4 KB | 表示 | |
XML形式データ | emd_22803_validation.xml.gz | 7.1 KB | 表示 | |
CIF形式データ | emd_22803_validation.cif.gz | 8.1 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-22803 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-22803 | 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_22803.map.gz / 形式: CCP4 / 大きさ: 282.6 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Replicative polymerase delta in complex with a primer/template and the PCNA clamp | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 0.826 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 : Pol delta-PCNA-DNA
+超分子 #1: Pol delta-PCNA-DNA
+超分子 #2: dsDNA
+超分子 #3: Proteins
+分子 #1: DNA (5'-D(P*AP*TP*GP*AP*CP*CP*AP*TP*GP*AP*TP*TP*AP*CP*GP*AP*AP*TP...
+分子 #2: DNA (25-MER)
+分子 #3: Proliferating cell nuclear antigen
+分子 #4: DNA polymerase
+分子 #5: POL31 isoform 1
+分子 #6: POL32 isoform 1
+分子 #7: MAGNESIUM ION
+分子 #8: IRON/SULFUR CLUSTER
+分子 #9: 2',3'-DIDEOXY-THYMIDINE-5'-TRIPHOSPHATE
+分子 #10: ZINC ION
-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
緩衝液 | pH: 7.5 |
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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: GATAN K3 (6k x 4k) / 平均電子線量: 68.0 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
最終 再構成 | 解像度のタイプ: BY AUTHOR / 解像度: 3.2 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 133468 |
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初期 角度割当 | タイプ: NOT APPLICABLE |
最終 角度割当 | タイプ: NOT APPLICABLE |