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データを開く
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基本情報
登録情報 | データベース: EMDB / ID: EMD-3979 | |||||||||
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タイトル | Post-catalytic P complex spliceosome with 3' splice site docked | |||||||||
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機能・相同性 | ![]() U2-type post-spliceosomal complex / spliceosomal complex disassembly / mRNA branch site recognition / U2-type post-mRNA release spliceosomal complex / cellular bud site selection / pre-mRNA 3'-splice site binding / post-mRNA release spliceosomal complex / nuclear mRNA surveillance / generation of catalytic spliceosome for first transesterification step / cis assembly of pre-catalytic spliceosome ...U2-type post-spliceosomal complex / spliceosomal complex disassembly / mRNA branch site recognition / U2-type post-mRNA release spliceosomal complex / cellular bud site selection / pre-mRNA 3'-splice site binding / post-mRNA release spliceosomal complex / nuclear mRNA surveillance / generation of catalytic spliceosome for first transesterification step / cis assembly of pre-catalytic spliceosome / splicing factor binding / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U4/U6 snRNP / 7-methylguanosine cap hypermethylation / pICln-Sm protein complex / U2-type catalytic step 1 spliceosome / pre-mRNA binding / snRNP binding / small nuclear ribonucleoprotein complex / SMN-Sm protein complex / spliceosomal tri-snRNP complex / U2-type spliceosomal complex / commitment complex / mRNA cis splicing, via spliceosome / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / U4 snRNP / U2 snRNP / U1 snRNP / poly(U) RNA binding / U2-type prespliceosome / precatalytic spliceosome / generation of catalytic spliceosome for second transesterification step / Formation of TC-NER Pre-Incision Complex / spliceosomal complex assembly / mRNA 5'-splice site recognition / Gap-filling DNA repair synthesis and ligation in TC-NER / mRNA 3'-splice site recognition / DNA replication origin binding / Dual incision in TC-NER / spliceosomal tri-snRNP complex assembly / Prp19 complex / protein K63-linked ubiquitination / U5 snRNA binding / DNA replication initiation / U5 snRNP / U2 snRNA binding / U6 snRNA binding / pre-mRNA intronic binding / spliceosomal snRNP assembly / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / positive regulation of cell cycle / catalytic step 2 spliceosome / nuclear periphery / RNA splicing / positive regulation of RNA splicing / spliceosomal complex / RING-type E3 ubiquitin transferase / mRNA splicing, via spliceosome / metallopeptidase activity / ubiquitin-protein transferase activity / ubiquitin protein ligase activity / RNA helicase activity / RNA helicase / DNA repair / GTPase activity / mRNA binding / chromatin binding / chromatin / GTP binding / ATP hydrolysis activity / mitochondrion / DNA binding / RNA binding / zinc ion binding / ATP binding / metal ion binding / identical protein binding / nucleus / cytosol / cytoplasm 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: 3.7 Å | |||||||||
![]() | Wilkinson ME / Fica SM / Galej WP / Norman CM / Newman AJ / Nagai K | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Postcatalytic spliceosome structure reveals mechanism of 3'-splice site selection. 著者: Max E Wilkinson / Sebastian M Fica / Wojciech P Galej / Christine M Norman / Andrew J Newman / Kiyoshi Nagai / ![]() 要旨: Introns are removed from eukaryotic messenger RNA precursors by the spliceosome in two transesterification reactions-branching and exon ligation. The mechanism of 3'-splice site recognition during ...Introns are removed from eukaryotic messenger RNA precursors by the spliceosome in two transesterification reactions-branching and exon ligation. The mechanism of 3'-splice site recognition during exon ligation has remained unclear. Here we present the 3.7-angstrom cryo-electron microscopy structure of the yeast P-complex spliceosome immediately after exon ligation. The 3'-splice site AG dinucleotide is recognized through non-Watson-Crick pairing with the 5' splice site and the branch-point adenosine. After the branching reaction, protein factors work together to remodel the spliceosome and stabilize a conformation competent for 3'-splice site docking, thereby promoting exon ligation. The structure accounts for the strict conservation of the GU and AG dinucleotides at the 5' and 3' ends of introns and provides insight into the catalytic mechanism of exon ligation. | |||||||||
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
-EMDBアーカイブ
マップデータ | ![]() | 390.9 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 63.2 KB 63.2 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 17 KB | 表示 | ![]() |
画像 | ![]() | 71 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
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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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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: 1.12 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 : P complex spliceosome with 3' exon docked
+超分子 #1: P complex spliceosome with 3' exon docked
+超分子 #2: P complex spliceosome
+超分子 #3: 3' exon
+分子 #1: U2 snRNA
+分子 #2: U5 snRNA
+分子 #3: U6 snRNA
+分子 #7: Ligated exons: UBC4 mRNA
+分子 #9: Intron lariat: UBC4 RNA
+分子 #4: Pre-mRNA-splicing factor Prp8
+分子 #5: Pre-mRNA-splicing factor SNU114
+分子 #6: Pre-mRNA-splicing factor CWC16
+分子 #8: Pre-mRNA-splicing factor CWC22
+分子 #10: Pre-mRNA-splicing factor PRP46
+分子 #11: Pre-mRNA-processing protein 45
+分子 #12: Pre-mRNA-splicing factor BUD31
+分子 #13: Pre-mRNA-splicing factor CWC2
+分子 #14: Pre-mRNA-splicing factor SLT11
+分子 #15: Pre-mRNA-splicing factor CEF1
+分子 #16: Pre-mRNA-splicing factor CWC15
+分子 #17: Pre-mRNA-splicing factor CWC21
+分子 #18: Pre-mRNA-splicing factor CLF1
+分子 #19: Pre-mRNA-splicing factor SYF1
+分子 #20: Pre-mRNA-splicing factor ATP-dependent RNA helicase PRP22
+分子 #21: U2 small nuclear ribonucleoprotein A'
+分子 #22: Unassigned structure
+分子 #23: U2 small nuclear ribonucleoprotein B''
+分子 #24: Pre-mRNA-splicing factor Prp18
+分子 #25: Small nuclear ribonucleoprotein-associated protein B
+分子 #26: Pre-mRNA-splicing factor SLU7
+分子 #27: Small nuclear ribonucleoprotein Sm D3
+分子 #28: Small nuclear ribonucleoprotein E
+分子 #29: Small nuclear ribonucleoprotein F
+分子 #30: Small nuclear ribonucleoprotein G
+分子 #31: Small nuclear ribonucleoprotein Sm D1
+分子 #32: Small nuclear ribonucleoprotein Sm D2
+分子 #33: Pre-mRNA-processing factor Prp17
+分子 #34: Pre-mRNA-splicing factor SNT309
+分子 #35: Pre-mRNA-processing factor Prp19
+分子 #36: Pre-mRNA-splicing factor SYF2
+分子 #37: MAGNESIUM ION
+分子 #38: INOSITOL HEXAKISPHOSPHATE
+分子 #39: GUANOSINE-5'-TRIPHOSPHATE
+分子 #40: 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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![]() | 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 | 1.4 mg/mL | ||||||||||||
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緩衝液 | pH: 7.9 構成要素:
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グリッド | モデル: Quantifoil R2/2 / 材質: COPPER / メッシュ: 400 / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: CONTINUOUS / 支持フィルム - Film thickness: 7.0 nm / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 雰囲気: AIR | ||||||||||||
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277 K / 装置: FEI VITROBOT MARK III 詳細: 3 uL sample was applied to the grid, left for 30s, then blotted for 3s and immediately plunged into liquid ethane.. |
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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 K2 SUMMIT (4k x 4k) 検出モード: COUNTING / デジタル化 - 画像ごとのフレーム数: 1-20 / 撮影したグリッド数: 1 / 実像数: 2295 / 平均露光時間: 12.0 sec. / 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 | C2レンズ絞り径: 50.0 µm / 最大 デフォーカス(補正後): 3.0 µm / 最小 デフォーカス(補正後): 0.2 µm / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm / 倍率(公称値): 105000 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |