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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-4658 | |||||||||
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タイトル | Structure of the human U5.U4/U6 tri-snRNP at 2.9A resolution. | |||||||||
![]() | Human U4.U5/U6 tri-snRNP, overall map. | |||||||||
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![]() | RNP complex / splicing / RNA / protein / spliceosome | |||||||||
機能・相同性 | ![]() Lsm2-8 complex / U6 snRNA 3'-end binding / spliceosomal snRNP complex / ribonucleoprotein complex localization / U4atac snRNP / positive regulation of cytotoxic T cell differentiation / maturation of 5S rRNA / RNA localization / U4atac snRNA binding / R-loop processing ...Lsm2-8 complex / U6 snRNA 3'-end binding / spliceosomal snRNP complex / ribonucleoprotein complex localization / U4atac snRNP / positive regulation of cytotoxic T cell differentiation / maturation of 5S rRNA / RNA localization / U4atac snRNA binding / R-loop processing / mRNA decay by 5' to 3' exoribonuclease / Lsm1-7-Pat1 complex / box C/D sno(s)RNA binding / U6 snRNP / PH domain binding / U2 snRNP binding / dense fibrillar component / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / histone pre-mRNA 3'end processing complex / cis assembly of pre-catalytic spliceosome / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / box C/D methylation guide snoRNP complex / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U4/U6 snRNP / protein methylation / U12-type spliceosomal complex / 7-methylguanosine cap hypermethylation / U1 snRNP binding / RNA splicing, via transesterification reactions / sno(s)RNA-containing ribonucleoprotein complex / methylosome / pICln-Sm protein complex / U2-type catalytic step 1 spliceosome / U4 snRNA binding / snRNP binding / small nuclear ribonucleoprotein complex / telomerase holoenzyme complex / SMN-Sm protein complex / P granule / telomerase RNA binding / spliceosomal tri-snRNP complex / U2-type precatalytic spliceosome / U2-type spliceosomal complex / commitment complex / mRNA cis splicing, via spliceosome / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / P-body assembly / U4 snRNP / U2 snRNP / RNA Polymerase II Transcription Termination / U3 snoRNA binding / U1 snRNP / U2-type prespliceosome / rRNA modification in the nucleus and cytosol / tRNA processing / precatalytic spliceosome / K63-linked polyubiquitin modification-dependent protein binding / spliceosomal complex assembly / mRNA catabolic process / mRNA Splicing - Minor Pathway / negative regulation of mRNA splicing, via spliceosome / nuclear-transcribed mRNA catabolic process / MLL1 complex / spliceosomal tri-snRNP complex assembly / protein deubiquitination / U5 snRNA binding / U5 snRNP / U2 snRNA binding / U6 snRNA binding / Major pathway of rRNA processing in the nucleolus and cytosol / pre-mRNA intronic binding / spliceosomal snRNP assembly / RNA processing / ribonucleoprotein complex binding / Cajal body / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / RNA splicing / maturation of SSU-rRNA / helicase activity / small-subunit processome / response to bacterium / spliceosomal complex / P-body / response to cocaine / mRNA splicing, via spliceosome / small GTPase binding / mRNA processing / osteoblast differentiation / cellular response to xenobiotic stimulus / cellular response to tumor necrosis factor / ATPase binding / ribosomal small subunit biogenesis / cellular response to lipopolysaccharide 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: 2.92 Å | |||||||||
![]() | Charenton C / Wilkinson ME | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Mechanism of 5' splice site transfer for human spliceosome activation. 著者: Clément Charenton / Max E Wilkinson / Kiyoshi Nagai / ![]() 要旨: The prespliceosome, comprising U1 and U2 small nuclear ribonucleoproteins (snRNPs) bound to the precursor messenger RNA 5' splice site (5'SS) and branch point sequence, associates with the U4/U6.U5 ...The prespliceosome, comprising U1 and U2 small nuclear ribonucleoproteins (snRNPs) bound to the precursor messenger RNA 5' splice site (5'SS) and branch point sequence, associates with the U4/U6.U5 tri-snRNP to form the fully assembled precatalytic pre-B spliceosome. Here, we report cryo-electron microscopy structures of the human pre-B complex captured before U1 snRNP dissociation at 3.3-angstrom core resolution and the human tri-snRNP at 2.9-angstrom resolution. U1 snRNP inserts the 5'SS-U1 snRNA helix between the two RecA domains of the Prp28 DEAD-box helicase. Adenosine 5'-triphosphate-dependent closure of the Prp28 RecA domains releases the 5'SS to pair with the nearby U6 ACAGAGA-box sequence presented as a mobile loop. The structures suggest that formation of the 5'SS-ACAGAGA helix triggers remodeling of an intricate protein-RNA network to induce Brr2 helicase relocation to its loading sequence in U4 snRNA, enabling Brr2 to unwind the U4/U6 snRNA duplex to allow U6 snRNA to form the catalytic center of the spliceosome. | |||||||||
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 | ![]() | 260.2 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 60.5 KB 60.5 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 14.8 KB | 表示 | ![]() |
画像 | ![]() | 181.4 KB | ||
Filedesc metadata | ![]() | 16.6 KB | ||
その他 | ![]() | 260.1 MB | ||
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 | ![]() 6qw6MC ![]() 4665C ![]() 4672C ![]() 4673C ![]() 4674C ![]() 4675C ![]() 4676C ![]() 4686C ![]() 4687C ![]() 4688C ![]() 4689C ![]() 4690C ![]() 6qx9C M: このマップから作成された原子モデル C: 同じ文献を引用 ( |
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類似構造データ | |
電子顕微鏡画像生データ | ![]() Data size: 2.3 TB Data #1: Dataset 1 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #2: Dataset 2 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #3: Dataset 3 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #4: Dataset 4 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #5: Dataset 5 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #6: Selected U4/U6.U5 tri-snRNP particles after Bayesian polishing [picked particles - single frame - processed] Data #7: Crude shifted preB particles [picked particles - single frame - unprocessed]) |
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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 | Human U4.U5/U6 tri-snRNP, overall map. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.022 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
-追加マップ: Composite map from the multi body refinement. For...
ファイル | emd_4658_additional.map | ||||||||||||
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注釈 | Composite map from the multi body refinement. For each map voxel, the map value was taken from the body with the highest local resolution as calculated with RELION. | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
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試料の構成要素
+全体 : Human U4/U6.U5 tri-snRNP
+超分子 #1: Human U4/U6.U5 tri-snRNP
+超分子 #2: U5.U4/U6 tri-snRNP
+超分子 #3: Probable ATP-dependent RNA helicase DDX23
+分子 #1: U4/U6.U5 small nuclear ribonucleoprotein 27 kDa protein
+分子 #3: Small nuclear ribonucleoprotein Sm D1
+分子 #4: Small nuclear ribonucleoprotein Sm D2
+分子 #5: Small nuclear ribonucleoprotein Sm D3
+分子 #6: U4/U6 small nuclear ribonucleoprotein Prp3
+分子 #7: U4/U6 small nuclear ribonucleoprotein Prp4
+分子 #8: U4/U6 small nuclear ribonucleoprotein Prp31
+分子 #9: NHP2-like protein 1
+分子 #10: Small nuclear ribonucleoprotein-associated proteins B and B'
+分子 #11: Small nuclear ribonucleoprotein E
+分子 #12: Small nuclear ribonucleoprotein F
+分子 #13: Small nuclear ribonucleoprotein G
+分子 #15: Pre-mRNA-processing-splicing factor 8
+分子 #16: U5 small nuclear ribonucleoprotein 200 kDa helicase
+分子 #17: 116 kDa U5 small nuclear ribonucleoprotein component
+分子 #18: Thioredoxin-like protein 4A
+分子 #19: Pre-mRNA-processing factor 6
+分子 #20: U5 small nuclear ribonucleoprotein 40 kDa protein
+分子 #21: Probable ATP-dependent RNA helicase DDX23
+分子 #23: U6 snRNA-associated Sm-like protein LSm2
+分子 #24: U6 snRNA-associated Sm-like protein LSm3
+分子 #25: U6 snRNA-associated Sm-like protein LSm4
+分子 #26: U6 snRNA-associated Sm-like protein LSm5
+分子 #27: U6 snRNA-associated Sm-like protein LSm6
+分子 #28: U6 snRNA-associated Sm-like protein LSm7
+分子 #29: U6 snRNA-associated Sm-like protein LSm8
+分子 #30: RNA-binding protein 42
+分子 #31: U4/U6.U5 tri-snRNP-associated protein 1
+分子 #32: U4/U6.U5 tri-snRNP-associated protein 2
+分子 #2: U4 snRNA
+分子 #14: U5 snRNA
+分子 #22: U6 snRNA
+分子 #33: INOSITOL HEXAKISPHOSPHATE
+分子 #34: MAGNESIUM ION
+分子 #35: GUANOSINE-5'-TRIPHOSPHATE
+分子 #36: 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 | 1.3 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: 300 / 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: 6 / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 時間: 20 sec. | ||||||||
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: Wait 30s, blot for 2s to 3s.. |
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電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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特殊光学系 | エネルギーフィルター - 名称: GIF Bioquantum / エネルギーフィルター - スリット幅: 20 eV |
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: 1-40 / 平均露光時間: 6.0 sec. / 平均電子線量: 50.0 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: 70.0 µm / 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm / 倍率(公称値): 130000 |
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 |