+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-4255 | |||||||||
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タイトル | human Bact spliceosome core structure | |||||||||
マップデータ | ||||||||||
試料 |
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機能・相同性 | 機能・相同性情報 RES complex / negative regulation of chemokine-mediated signaling pathway / snoRNA splicing / U11/U12 snRNP / regulation of retinoic acid receptor signaling pathway / post-mRNA release spliceosomal complex / B-WICH complex / regulation of vitamin D receptor signaling pathway / splicing factor binding / U12-type spliceosomal complex ...RES complex / negative regulation of chemokine-mediated signaling pathway / snoRNA splicing / U11/U12 snRNP / regulation of retinoic acid receptor signaling pathway / post-mRNA release spliceosomal complex / B-WICH complex / regulation of vitamin D receptor signaling pathway / splicing factor binding / U12-type spliceosomal complex / embryonic brain development / nuclear retinoic acid receptor binding / positive regulation of androgen receptor activity / Prp19 complex / mRNA 3'-end processing / blastocyst formation / U2-type catalytic step 1 spliceosome / RNA splicing, via transesterification reactions / pre-mRNA binding / regulation of mRNA splicing, via spliceosome / C2H2 zinc finger domain binding / U2-type spliceosomal complex / U2-type precatalytic spliceosome / positive regulation by host of viral transcription / mRNA cis splicing, via spliceosome / positive regulation of vitamin D receptor signaling pathway / U2-type prespliceosome assembly / nuclear vitamin D receptor binding / Transport of Mature mRNA derived from an Intron-Containing Transcript / U2-type catalytic step 2 spliceosome / Notch binding / Regulation of gene expression in late stage (branching morphogenesis) pancreatic bud precursor cells / positive regulation of mRNA splicing, via spliceosome / RUNX3 regulates NOTCH signaling / U2 snRNP / RHOBTB1 GTPase cycle / SAGA complex / NOTCH4 Intracellular Domain Regulates Transcription / RNA Polymerase II Transcription Termination / positive regulation of transcription by RNA polymerase III / NOTCH3 Intracellular Domain Regulates Transcription / WD40-repeat domain binding / positive regulation of neurogenesis / U2-type prespliceosome / nuclear androgen receptor binding / precatalytic spliceosome / K63-linked polyubiquitin modification-dependent protein binding / cyclosporin A binding / positive regulation of transcription by RNA polymerase I / Notch-HLH transcription pathway / positive regulation of transforming growth factor beta receptor signaling pathway / Formation of paraxial mesoderm / mRNA Splicing - Minor Pathway / spliceosomal complex assembly / SMAD binding / regulation of RNA splicing / protein peptidyl-prolyl isomerization / mRNA 3'-splice site recognition / protein localization to nucleus / spliceosomal tri-snRNP complex assembly / intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / U5 snRNA binding / U5 snRNP / RHOBTB2 GTPase cycle / positive regulation of G1/S transition of mitotic cell cycle / retinoic acid receptor signaling pathway / U2 snRNA binding / U6 snRNA binding / regulation of DNA repair / Cajal body / pre-mRNA intronic binding / cellular response to retinoic acid / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / RNA splicing / positive regulation of RNA splicing / positive regulation of protein export from nucleus / nuclear receptor coactivator activity / DNA damage checkpoint signaling / response to cocaine / peptidylprolyl isomerase / nuclear receptor binding / stem cell differentiation / peptidyl-prolyl cis-trans isomerase activity / RNA polymerase II transcription regulatory region sequence-specific DNA binding / Downregulation of SMAD2/3:SMAD4 transcriptional activity / spliceosomal complex / RING-type E3 ubiquitin transferase / B-WICH complex positively regulates rRNA expression / negative regulation of protein catabolic process / NOTCH1 Intracellular Domain Regulates Transcription / mRNA splicing, via spliceosome / fibrillar center / Pre-NOTCH Transcription and Translation / positive regulation of neuron projection development / Constitutive Signaling by NOTCH1 PEST Domain Mutants / Constitutive Signaling by NOTCH1 HD+PEST Domain Mutants / nuclear matrix 類似検索 - 分子機能 | |||||||||
生物種 | Homo sapiens (ヒト) / Human (ヒト) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.4 Å | |||||||||
データ登録者 | Haselbach D / Komarov I / Agafonov D / Hartmuth K / Graf B / Kastner B / Luehrmann R / Stark H | |||||||||
引用 | ジャーナル: Cell / 年: 2018 タイトル: Structure and Conformational Dynamics of the Human Spliceosomal B Complex. 著者: David Haselbach / Ilya Komarov / Dmitry E Agafonov / Klaus Hartmuth / Benjamin Graf / Olexandr Dybkov / Henning Urlaub / Berthold Kastner / Reinhard Lührmann / Holger Stark / 要旨: The spliceosome is a highly dynamic macromolecular complex that precisely excises introns from pre-mRNA. Here we report the cryo-EM 3D structure of the human B spliceosome at 3.4 Å resolution. In ...The spliceosome is a highly dynamic macromolecular complex that precisely excises introns from pre-mRNA. Here we report the cryo-EM 3D structure of the human B spliceosome at 3.4 Å resolution. In the B state, the spliceosome is activated but not catalytically primed, so that it is functionally blocked prior to the first catalytic step of splicing. The spliceosomal core is similar to the yeast B spliceosome; important differences include the presence of the RNA helicase aquarius and peptidyl prolyl isomerases. To examine the overall dynamic behavior of the purified spliceosome, we developed a principal component analysis-based approach. Calculating the energy landscape revealed eight major conformational states, which we refined to higher resolution. Conformational differences of the highly flexible structural components between these eight states reveal how spliceosomal components contribute to the assembly of the spliceosome, allowing it to generate a dynamic interaction network required for its subsequent catalytic activation. | |||||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | EMマップ: SurfViewMolmilJmol/JSmol |
添付画像 |
-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_4255.map.gz | 25.1 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-4255-v30.xml emd-4255.xml | 54.1 KB 54.1 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_4255.png | 53.1 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-4255 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-4255 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_4255_validation.pdf.gz | 233.7 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_4255_full_validation.pdf.gz | 232.8 KB | 表示 | |
XML形式データ | emd_4255_validation.xml.gz | 7.2 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4255 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4255 | HTTPS FTP |
-関連構造データ
関連構造データ | 6ff4MC 4233C 4234C 4235C 4236C 4237C 4238C 4239C 4240C 4247C 4248C 4249C 4250C 4251C 4252C 4253C 4254C 6ff7C C: 同じ文献を引用 (文献) 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 |
-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_4255.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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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: 1.16 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
-試料の構成要素
+全体 : human Bact spliceosome state 1 unmasked
+超分子 #1: human Bact spliceosome state 1 unmasked
+分子 #1: RNA-binding motif protein, X-linked 2
+分子 #3: BUD13 homolog
+分子 #6: Splicing factor 3A subunit 2
+分子 #7: Splicing factor 3B subunit 2
+分子 #8: Pre-mRNA-processing-splicing factor 8
+分子 #9: 116 kDa U5 small nuclear ribonucleoprotein component
+分子 #10: SNW domain-containing protein 1
+分子 #11: Pleiotropic regulator 1
+分子 #12: Pre-mRNA-processing factor 17
+分子 #13: Cell division cycle 5-like protein
+分子 #14: Crooked neck-like protein 1
+分子 #15: Pre-mRNA-splicing factor RBM22
+分子 #16: Protein BUD31 homolog
+分子 #17: Spliceosome-associated protein CWC15 homolog
+分子 #18: Serine/arginine repetitive matrix protein 2
+分子 #19: Peptidyl-prolyl cis-trans isomerase-like 1
+分子 #20: Serine/arginine repetitive matrix protein 1
+分子 #22: Peptidyl-prolyl cis-trans isomerase CWC27 homolog
+分子 #23: RING finger protein 113A
+分子 #24: Splicing factor 3B subunit 1
+分子 #25: Splicing factor 3B subunit 3
+分子 #26: Splicing factor 3B subunit 5
+分子 #27: PHD finger-like domain-containing protein 5A
+分子 #28: Splicing factor 3B subunit 6
+分子 #2: U2 snRNA
+分子 #4: U5 snRNA
+分子 #5: U6 snRNA
+分子 #21: pre mRNA
+分子 #29: MAGNESIUM ION
+分子 #30: INOSITOL HEXAKISPHOSPHATE
+分子 #31: GUANOSINE-5'-TRIPHOSPHATE
+分子 #32: 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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解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
濃度 | 0.05 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 R3.5/1 / 材質: COPPER / メッシュ: 400 / 支持フィルム - #0 - Film type ID: 1 / 支持フィルム - #0 - 材質: CARBON / 支持フィルム - #0 - トポロジー: HOLEY / 支持フィルム - #1 - Film type ID: 2 / 支持フィルム - #1 - 材質: CARBON / 支持フィルム - #1 - トポロジー: CONTINUOUS | ||||||||||||
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 75 % / チャンバー内温度: 277 K / 装置: LEICA EM GP / 詳細: blot with blotting sensor. |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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アライメント法 | Coma free - Residual tilt: 14.0 mrad |
特殊光学系 | 球面収差補正装置: Microscope was modified with a Cs corrector with two hexapoles elements |
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 / デジタル化 - サンプリング間隔: 14.0 µm / 撮影したグリッド数: 1 / 実像数: 32000 / 平均露光時間: 1.0 sec. / 平均電子線量: 40.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | C2レンズ絞り径: 70.0 µm / 照射モード: SPOT SCAN / 撮影モード: BRIGHT FIELD / Cs: 0.001 mm / 最大 デフォーカス(公称値): 4.5 µm / 最小 デフォーカス(公称値): 0.8 µm / 倍率(公称値): 59000 |
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 |
+画像解析
-原子モデル構築 1
精密化 | プロトコル: RIGID BODY FIT |
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得られたモデル | PDB-6ff4: |