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基本情報
登録情報 | データベース: EMDB / ID: EMD-7851 | |||||||||
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タイトル | Cryo-EM structure of RAG in complex with melted 12-RSS and unmelted 23-RSS substrates | |||||||||
![]() | RAG in complex with 12-RSS and 23-RSS substrate DNAs | |||||||||
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![]() | V(D)J recombination / RAG complex / Melted RSS / Unmelted RSS / Recombination-DNA complex | |||||||||
機能・相同性 | ![]() somatic diversification of immune receptors via germline recombination within a single locus / hematopoietic or lymphoid organ development / DNA recombinase complex / endodeoxyribonuclease complex / protein-DNA complex assembly / lymphocyte differentiation / immunoglobulin V(D)J recombination / V(D)J recombination / phosphatidylinositol-3,4-bisphosphate binding / phosphatidylinositol-3,5-bisphosphate binding ...somatic diversification of immune receptors via germline recombination within a single locus / hematopoietic or lymphoid organ development / DNA recombinase complex / endodeoxyribonuclease complex / protein-DNA complex assembly / lymphocyte differentiation / immunoglobulin V(D)J recombination / V(D)J recombination / phosphatidylinositol-3,4-bisphosphate binding / phosphatidylinositol-3,5-bisphosphate binding / detection of maltose stimulus / maltose transport complex / phosphatidylinositol-3,4,5-trisphosphate binding / carbohydrate transport / T cell differentiation / carbohydrate transmembrane transporter activity / maltose binding / maltose transport / maltodextrin transmembrane transport / ATP-binding cassette (ABC) transporter complex, substrate-binding subunit-containing / : / phosphatidylinositol-4,5-bisphosphate binding / phosphatidylinositol binding / B cell differentiation / ATP-binding cassette (ABC) transporter complex / thymus development / cell chemotaxis / RING-type E3 ubiquitin transferase / ubiquitin-protein transferase activity / ubiquitin protein ligase activity / chromatin organization / T cell differentiation in thymus / outer membrane-bounded periplasmic space / endonuclease activity / DNA recombination / adaptive immune response / sequence-specific DNA binding / histone binding / 加水分解酵素; エステル加水分解酵素 / periplasmic space / DNA damage response / chromatin binding / magnesium ion binding / protein homodimerization activity / DNA binding / zinc ion binding / metal ion binding / nucleus / membrane 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: 4.29 Å | |||||||||
![]() | Wu H / Liao M | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: DNA melting initiates the RAG catalytic pathway. 著者: Heng Ru / Wei Mi / Pengfei Zhang / Frederick W Alt / David G Schatz / Maofu Liao / Hao Wu / ![]() 要旨: The mechanism for initiating DNA cleavage by DDE-family enzymes, including the RAG endonuclease, which initiates V(D)J recombination, is not well understood. Here we report six cryo-EM structures of ...The mechanism for initiating DNA cleavage by DDE-family enzymes, including the RAG endonuclease, which initiates V(D)J recombination, is not well understood. Here we report six cryo-EM structures of zebrafish RAG in complex with one or two intact recombination signal sequences (RSSs), at up to 3.9-Å resolution. Unexpectedly, these structures reveal DNA melting at the heptamer of the RSSs, thus resulting in a corkscrew-like rotation of coding-flank DNA and the positioning of the scissile phosphate in the active site. Substrate binding is associated with dimer opening and a piston-like movement in RAG1, first outward to accommodate unmelted DNA and then inward to wedge melted DNA. These precleavage complexes show limited base-specific contacts of RAG at the conserved terminal CAC/GTG sequence of the heptamer, thus suggesting conservation based on a propensity to unwind. CA and TG overwhelmingly dominate terminal sequences in transposons and retrotransposons, thereby implicating a universal mechanism for DNA melting during the initiation of retroviral integration and DNA transposition. | |||||||||
履歴 |
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構造の表示
ムービー |
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構造ビューア | EMマップ: ![]() ![]() ![]() |
添付画像 |
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ダウンロードとリンク
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マップデータ | ![]() | 22.3 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 22 KB 22 KB | 表示 表示 | ![]() |
画像 | ![]() | 64.4 KB | ||
Filedesc metadata | ![]() | 7.1 KB | ||
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 | ![]() 6dbvMC ![]() 7843C ![]() 7844C ![]() 7845C ![]() 7846C ![]() 7847C ![]() 7848C ![]() 7849C ![]() 7850C ![]() 7852C ![]() 7853C ![]() 6dbiC ![]() 6dbjC ![]() 6dblC ![]() 6dboC ![]() 6dbqC ![]() 6dbrC ![]() 6dbtC ![]() 6dbuC ![]() 6dbwC ![]() 6dbxC 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 |
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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 | RAG in complex with 12-RSS and 23-RSS substrate DNAs | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.238 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
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試料の構成要素
+全体 : RAG in complex with 12-RSS and 23-RSS substrate DNAs
+超分子 #1: RAG in complex with 12-RSS and 23-RSS substrate DNAs
+分子 #1: Recombination activating gene 1 - MBP chimera
+分子 #2: Recombination activating gene 2
+分子 #3: Recombination activating gene 1 - MBP chimera
+分子 #4: Forward strand of 12-RSS substrate DNA
+分子 #5: Reverse strand of 12-RSS substrate DNA
+分子 #6: Forward strand of 23-RSS substrate DNA
+分子 #7: Reverse strand of 23-RSS substrate DNA
+分子 #8: ZINC ION
+分子 #9: CALCIUM 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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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
緩衝液 | pH: 7.5 構成要素:
詳細: Solutions were made fresh from concentrated to avoid microbial contamination. | |||||||||||||||
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凍結 | 凍結剤: ETHANE | |||||||||||||||
詳細 | This sample was monodisperse. |
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電子顕微鏡法
顕微鏡 | FEI POLARA 300 |
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撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 平均電子線量: 47.0 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 |
実験機器 | ![]() モデル: Tecnai Polara / 画像提供: FEI Company |
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画像解析
初期モデル | モデルのタイプ: NONE |
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最終 再構成 | 想定した対称性 - 点群: C1 (非対称) / 解像度のタイプ: BY AUTHOR / 解像度: 4.29 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 45159 |
初期 角度割当 | タイプ: PROJECTION MATCHING |
最終 角度割当 | タイプ: PROJECTION MATCHING |