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基本情報
登録情報 | データベース: EMDB / ID: EMD-11696 | |||||||||
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タイトル | Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion) | |||||||||
![]() | Masked/sharpened map of SF3b/U2 snRNP region of pre-Bact-2 spliceosome. | |||||||||
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![]() | Complex / spliceosome / catalytic activation / splicing | |||||||||
機能・相同性 | ![]() U11/U12 snRNP / B-WICH complex / splicing factor binding / U12-type spliceosomal complex / miRNA processing / Prp19 complex / mRNA 3'-end processing / blastocyst formation / RNA splicing, via transesterification reactions / regulation of mRNA splicing, via spliceosome ...U11/U12 snRNP / B-WICH complex / splicing factor binding / U12-type spliceosomal complex / miRNA processing / Prp19 complex / mRNA 3'-end processing / blastocyst formation / RNA splicing, via transesterification reactions / regulation of mRNA splicing, via spliceosome / U2-type spliceosomal complex / U2-type precatalytic spliceosome / Transport of Mature mRNA derived from an Intron-Containing Transcript / transcription regulator inhibitor activity / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / U2 snRNP / SAGA complex / positive regulation of mRNA splicing, via spliceosome / RNA Polymerase II Transcription Termination / RHOBTB1 GTPase cycle / positive regulation of transcription by RNA polymerase III / WD40-repeat domain binding / U2-type prespliceosome / positive regulation of transcription by RNA polymerase I / precatalytic spliceosome / spliceosomal complex assembly / mRNA Splicing - Minor Pathway / regulation of RNA splicing / mRNA 3'-splice site recognition / localization / RHOBTB2 GTPase cycle / Protein methylation / U2 snRNA binding / regulation of DNA repair / negative regulation of canonical NF-kappaB signal transduction / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / RNA splicing / DNA damage checkpoint signaling / stem cell differentiation / RNA polymerase II transcription regulatory region sequence-specific DNA binding / spliceosomal complex / B-WICH complex positively regulates rRNA expression / negative regulation of protein catabolic process / mRNA processing / nuclear matrix / positive regulation of neuron projection development / mRNA splicing, via spliceosome / double-stranded DNA binding / DNA-binding transcription activator activity, RNA polymerase II-specific / nuclear membrane / DNA recombination / DNA replication / DNA-binding transcription factor activity, RNA polymerase II-specific / nuclear speck / chromatin remodeling / cell cycle / intracellular membrane-bounded organelle / DNA repair / mRNA binding / DNA damage response / protein-containing complex binding / nucleolus / regulation of transcription by RNA polymerase II / positive regulation of DNA-templated transcription / endoplasmic reticulum / positive regulation of transcription by RNA polymerase II / protein-containing complex / DNA binding / RNA binding / zinc ion binding / nucleoplasm / identical protein binding / membrane / nucleus / metal ion binding / cytoplasm / cytosol 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: 4.5 Å | |||||||||
![]() | Townsend C / Kastner B | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Mechanism of protein-guided folding of the active site U2/U6 RNA during spliceosome activation. 著者: Cole Townsend / Majety N Leelaram / Dmitry E Agafonov / Olexandr Dybkov / Cindy L Will / Karl Bertram / Henning Urlaub / Berthold Kastner / Holger Stark / Reinhard Lührmann / ![]() 要旨: Spliceosome activation involves extensive protein and RNA rearrangements that lead to formation of a catalytically active U2/U6 RNA structure. At present, little is known about the assembly pathway ...Spliceosome activation involves extensive protein and RNA rearrangements that lead to formation of a catalytically active U2/U6 RNA structure. At present, little is known about the assembly pathway of the latter and the mechanism whereby proteins aid its proper folding. Here, we report the cryo-electron microscopy structures of two human, activated spliceosome precursors (that is, pre-B complexes) at core resolutions of 3.9 and 4.2 angstroms. These structures elucidate the order of the numerous protein exchanges that occur during activation, the mutually exclusive interactions that ensure the correct order of ribonucleoprotein rearrangements needed to form the U2/U6 catalytic RNA, and the stepwise folding pathway of the latter. Structural comparisons with mature B complexes reveal the molecular mechanism whereby a conformational change in the scaffold protein PRP8 facilitates final three-dimensional folding of the U2/U6 catalytic RNA. | |||||||||
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: ![]() ![]() ![]() |
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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 | ![]() | 12.2 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 34 KB 34 KB | 表示 表示 | ![]() |
画像 | ![]() | 25 KB | ||
Filedesc metadata | ![]() | 11.3 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 349.4 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 349 KB | 表示 | |
XML形式データ | ![]() | 7 KB | 表示 | |
CIF形式データ | ![]() | 8 KB | 表示 | |
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 | ![]() 7abhMC ![]() 7aavC ![]() 7abfC ![]() 7abgC ![]() 7abiC 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 | |
電子顕微鏡画像生データ | ![]() Data #1: Motion-corrected micrographs (without dose-weighting) of human pre-Bact spliceosome [micrographs - single frame] Data #2: Motion-corrected micrographs (with dose-weighting) of human pre-Bact spliceosome [micrographs - single frame]) |
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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 | Masked/sharpened map of SF3b/U2 snRNP region of pre-Bact-2 spliceosome. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.16 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 : Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)
+超分子 #1: Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)
+超分子 #2: Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)
+超分子 #3: MINX M3 pre-mRNA
+分子 #1: Cell division cycle 5-like protein
+分子 #2: PHD finger-like domain-containing protein 5A
+分子 #4: Splicing factor 3A subunit 2
+分子 #5: Splicing factor 3A subunit 3
+分子 #6: Splicing factor 3B subunit 1
+分子 #7: Splicing factor 3B subunit 2
+分子 #8: Splicing factor 3B subunit 3
+分子 #9: Splicing factor 3B subunit 4
+分子 #10: Splicing factor 3B subunit 5
+分子 #11: Splicing factor 3B subunit 6
+分子 #12: Smad nuclear-interacting protein 1
+分子 #13: RNA-binding motif protein, X-linked 2
+分子 #14: Serine/arginine repetitive matrix protein 1
+分子 #16: DNA/RNA-binding protein KIN17
+分子 #3: MINX M3 pre-mRNA
+分子 #15: U2 snRNA
-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: 7.9 |
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グリッド | モデル: Quantifoil R3.5/1 / 材質: COPPER / 支持フィルム - 材質: CARBON / 支持フィルム - トポロジー: CONTINUOUS |
凍結 | 凍結剤: ETHANE / 装置: FEI VITROBOT MARK IV |
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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: FEI FALCON III (4k x 4k) 検出モード: INTEGRATING / デジタル化 - サイズ - 横: 4096 pixel / デジタル化 - サイズ - 縦: 4096 pixel / 平均露光時間: 1.0 sec. / 平均電子線量: 2.25 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: SPOT SCAN / 撮影モード: BRIGHT FIELD |
実験機器 | ![]() モデル: Titan Krios / 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: OTHER / 詳細: cryoSPARC ab initio reconstruction |
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最終 再構成 | 解像度のタイプ: BY AUTHOR / 解像度: 4.5 Å / 解像度の算出法: FSC 0.143 CUT-OFF / ソフトウェア - 名称: RELION (ver. 3.0) / 使用した粒子像数: 39336 |
初期 角度割当 | タイプ: MAXIMUM LIKELIHOOD |
最終 角度割当 | タイプ: MAXIMUM LIKELIHOOD |
-原子モデル構築 1
精密化 | 空間: REAL / プロトコル: RIGID BODY FIT |
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得られたモデル | ![]() PDB-7abh: |