+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-29415 | ||||||||||||
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タイトル | EM map of S. cerevisiae Rad24-RFC loading the 9-1-1 clamp onto a 10-nt gapped DNA in step 4 (partially closed 9-1-1 and stably bound chamber DNA) | ||||||||||||
マップデータ | EM map of S. cerevisiae Rad24-RFC loading the 9-1-1 clamp onto a 10-nt gapped DNA in step 4 (partially closed 9-1-1 and stably bound chamber DNA) | ||||||||||||
試料 |
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キーワード | DNA damage repair / Rad24-RFC / 9-1-1 clamp / DNA clamp / alternative clamp loader / DNA damage signaling / DNA BINDING PROTEIN-DNA complex / CELL CYCLE-DNA complex | ||||||||||||
機能・相同性 | 機能・相同性情報 meiotic DNA integrity checkpoint signaling / checkpoint clamp complex / DNA clamp unloading / Rad17 RFC-like complex / Gap-filling DNA repair synthesis and ligation in GG-NER / Elg1 RFC-like complex / Ctf18 RFC-like complex / DNA replication factor C complex / Polymerase switching / DNA clamp loader activity ...meiotic DNA integrity checkpoint signaling / checkpoint clamp complex / DNA clamp unloading / Rad17 RFC-like complex / Gap-filling DNA repair synthesis and ligation in GG-NER / Elg1 RFC-like complex / Ctf18 RFC-like complex / DNA replication factor C complex / Polymerase switching / DNA clamp loader activity / Translesion Synthesis by POLH / telomere maintenance via recombination / Translesion synthesis by REV1 / Translesion synthesis by POLK / Translesion synthesis by POLI / DNA replication checkpoint signaling / Activation of ATR in response to replication stress / Termination of translesion DNA synthesis / mitotic DNA replication checkpoint signaling / reciprocal meiotic recombination / mitotic intra-S DNA damage checkpoint signaling / recombinational repair / sister chromatid cohesion / mitotic sister chromatid cohesion / leading strand elongation / Gap-filling DNA repair synthesis and ligation in TC-NER / Dual incision in TC-NER / subtelomeric heterochromatin formation / mismatch repair / 3'-5' exonuclease activity / telomere maintenance / DNA damage checkpoint signaling / meiotic cell cycle / cellular response to ionizing radiation / nucleotide-excision repair / double-strand break repair via homologous recombination / DNA-templated DNA replication / double-strand break repair / site of double-strand break / double-stranded DNA binding / damaged DNA binding / chromosome, telomeric region / DNA repair / chromatin binding / ATP hydrolysis activity / DNA binding / ATP binding / nucleus / cytosol 類似検索 - 分子機能 | ||||||||||||
生物種 | synthetic construct (人工物) / 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: 2.9 Å | ||||||||||||
データ登録者 | Zheng F / Georgescu R / Yao YN / O'Donnell ME / Li H | ||||||||||||
資金援助 | 米国, 3件
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引用 | ジャーナル: bioRxiv / 年: 2023 タイトル: Structures of 9-1-1 DNA checkpoint clamp loading at gaps from start to finish and ramification to biology. 著者: Fengwei Zheng / Roxana E Georgescu / Nina Y Yao / Michael E O'Donnell / Huilin Li / 要旨: Recent structural studies show the Rad24-RFC loads the 9-1-1 checkpoint clamp onto a recessed 5' end by binding the 5' DNA on Rad24 at an external surface site and threading the 3' ssDNA into the ...Recent structural studies show the Rad24-RFC loads the 9-1-1 checkpoint clamp onto a recessed 5' end by binding the 5' DNA on Rad24 at an external surface site and threading the 3' ssDNA into the well-established internal chamber and into 9-1-1. We find here that Rad24-RFC loads 9-1-1 onto DNA gaps in preference to a recessed 5' DNA end, thus presumably leaving 9-1-1 on a 3' ss/ds DNA after Rad24-RFC ejects from the 5' gap end and may explain reports of 9-1-1 directly functioning in DNA repair with various TLS polymerases, in addition to signaling the ATR kinase. To gain a deeper understanding of 9-1-1 loading at gaps we report high-resolution structures of Rad24-RFC during loading of 9-1-1 onto 10-nt and 5-nt gapped DNAs. At a 10-nt gap we captured five Rad24-RFC-9-1-1 loading intermediates in which the 9-1-1 DNA entry gate varies from fully open to fully closed around DNA using ATPγS, supporting the emerging view that ATP hydrolysis is not needed for clamp opening/closing, but instead for dissociation of the loader from the clamp encircling DNA. The structure of Rad24-RFC-9-1-1 at a 5-nt gap shows a 180° axially rotated 3'-dsDNA which orients the template strand to bridge the 3'- and 5'- junctions with a minimum 5-nt ssDNA. The structures reveal a unique loop on Rad24 that limits the length of dsDNA in the inner chamber, and inability to melt DNA ends unlike RFC, thereby explaining Rad24-RFC's preference for a preexisting ssDNA gap and suggesting a direct role in gap repair in addition to its checkpoint role. | ||||||||||||
履歴 |
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-構造の表示
添付画像 |
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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 | emd_29415.map.gz | 266.8 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-29415-v30.xml emd-29415.xml | 28.9 KB 28.9 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_29415_fsc.xml | 13.9 KB | 表示 | FSCデータファイル |
画像 | emd_29415.png | 72.1 KB | ||
Filedesc metadata | emd-29415.cif.gz | 8.7 KB | ||
その他 | emd_29415_half_map_1.map.gz emd_29415_half_map_2.map.gz | 262.6 MB 262.6 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-29415 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-29415 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_29415_validation.pdf.gz | 1.2 MB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_29415_full_validation.pdf.gz | 1.2 MB | 表示 | |
XML形式データ | emd_29415_validation.xml.gz | 22.9 KB | 表示 | |
CIF形式データ | emd_29415_validation.cif.gz | 29.7 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-29415 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-29415 | 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_29415.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 | EM map of S. cerevisiae Rad24-RFC loading the 9-1-1 clamp onto a 10-nt gapped DNA in step 4 (partially closed 9-1-1 and stably bound chamber DNA) | ||||||||||||||||||||||||||||||||||||
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: 0.828 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
-ハーフマップ: half map A
ファイル | emd_29415_half_map_1.map | ||||||||||||
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注釈 | half map 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: half map B
ファイル | emd_29415_half_map_2.map | ||||||||||||
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注釈 | half map B | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-試料の構成要素
+全体 : Rad24-RFC-911 clamp-DNA
+超分子 #1: Rad24-RFC-911 clamp-DNA
+超分子 #2: DNA
+超分子 #3: Proteins complex
+分子 #1: Checkpoint protein RAD24
+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: DNA damage checkpoint control protein MEC3
+分子 #7: DNA damage checkpoint control protein RAD17
+分子 #8: DDC1 isoform 1
+分子 #9: Template strand
+分子 #10: Primer strand 1
+分子 #11: Primer strand 2
+分子 #12: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER
+分子 #13: MAGNESIUM ION
+分子 #14: ADENOSINE-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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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: 64.0 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 最大 デフォーカス(公称値): 1.9000000000000001 µm 最小 デフォーカス(公称値): 1.3 µm |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |