+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-3545 | |||||||||
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タイトル | Cryo-EM structure of a human spliceosome activated for step 2 of splicing (C* complex) | |||||||||
マップデータ | None | |||||||||
試料 |
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機能・相同性 | 機能・相同性情報 negative regulation of selenocysteine incorporation / regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / cellular response to selenite ion / spliceosomal complex disassembly / selenocysteine insertion sequence binding / exon-exon junction complex / regulation of translation at postsynapse, modulating synaptic transmission / regulation of retinoic acid receptor signaling pathway / post-mRNA release spliceosomal complex / U2 snRNP binding ...negative regulation of selenocysteine incorporation / regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / cellular response to selenite ion / spliceosomal complex disassembly / selenocysteine insertion sequence binding / exon-exon junction complex / regulation of translation at postsynapse, modulating synaptic transmission / regulation of retinoic acid receptor signaling pathway / post-mRNA release spliceosomal complex / U2 snRNP binding / U7 snRNA binding / negative regulation of excitatory postsynaptic potential / histone pre-mRNA DCP binding / 3'-5' RNA helicase activity / U7 snRNP / generation of catalytic spliceosome for first transesterification step / Z-decay: degradation of maternal mRNAs by zygotically expressed factors / regulation of vitamin D receptor signaling pathway / histone pre-mRNA 3'end processing complex / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / Deadenylation of mRNA / embryonic cranial skeleton morphogenesis / protein methylation / U12-type spliceosomal complex / embryonic brain development / methylosome / nuclear retinoic acid receptor binding / 7-methylguanosine cap hypermethylation / positive regulation of androgen receptor activity / M-decay: degradation of maternal mRNAs by maternally stored factors / Prp19 complex / poly(A) binding / U1 snRNP binding / pICln-Sm protein complex / mRNA 3'-end processing / ATP-dependent activity, acting on RNA / sno(s)RNA-containing ribonucleoprotein complex / snRNP binding / U2-type catalytic step 1 spliceosome / RNA splicing, via transesterification reactions / small nuclear ribonucleoprotein complex / pre-mRNA binding / regulation of mRNA splicing, via spliceosome / C2H2 zinc finger domain binding / SMN-Sm protein complex / telomerase RNA binding / spliceosomal tri-snRNP complex / telomerase holoenzyme complex / P granule / 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 / commitment complex / 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 / U4 snRNP / positive regulation of mRNA splicing, via spliceosome / RUNX3 regulates NOTCH signaling / U2 snRNP / NOTCH4 Intracellular Domain Regulates Transcription / RNA Polymerase II Transcription Termination / U1 snRNP / exploration behavior / ubiquitin-ubiquitin ligase activity / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / NOTCH3 Intracellular Domain Regulates Transcription / WD40-repeat domain binding / positive regulation of neurogenesis / U2-type prespliceosome / lipid biosynthetic process / nuclear androgen receptor binding / precatalytic spliceosome / K63-linked polyubiquitin modification-dependent protein binding / cyclosporin A binding / 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 / mitotic G2 DNA damage checkpoint signaling / associative learning / protein peptidyl-prolyl isomerization / protein K63-linked ubiquitination / blastocyst development / protein localization to nucleus / spliceosomal tri-snRNP complex assembly / intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / embryonic organ development / transcription-coupled nucleotide-excision repair / U5 snRNA binding / U5 snRNP / positive regulation of G1/S transition of mitotic cell cycle 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: 5.9 Å | |||||||||
データ登録者 | Bertram K / Liu WT | |||||||||
引用 | ジャーナル: Nature / 年: 2017 タイトル: Cryo-EM structure of a human spliceosome activated for step 2 of splicing. 著者: Karl Bertram / Dmitry E Agafonov / Wen-Ti Liu / Olexandr Dybkov / Cindy L Will / Klaus Hartmuth / Henning Urlaub / Berthold Kastner / Holger Stark / Reinhard Lührmann / 要旨: Spliceosome rearrangements facilitated by RNA helicase PRP16 before catalytic step two of splicing are poorly understood. Here we report a 3D cryo-electron microscopy structure of the human ...Spliceosome rearrangements facilitated by RNA helicase PRP16 before catalytic step two of splicing are poorly understood. Here we report a 3D cryo-electron microscopy structure of the human spliceosomal C complex stalled directly after PRP16 action (C*). The architecture of the catalytic U2-U6 ribonucleoprotein (RNP) core of the human C* spliceosome is very similar to that of the yeast pre-Prp16 C complex. However, in C* the branched intron region is separated from the catalytic centre by approximately 20 Å, and its position close to the U6 small nuclear RNA ACAGA box is stabilized by interactions with the PRP8 RNase H-like and PRP17 WD40 domains. RNA helicase PRP22 is located about 100 Å from the catalytic centre, suggesting that it destabilizes the spliced mRNA after step two from a distance. Comparison of the structure of the yeast C and human C* complexes reveals numerous RNP rearrangements that are likely to be facilitated by PRP16, including a large-scale movement of the U2 small nuclear RNP. | |||||||||
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_3545.map.gz | 164.8 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-3545-v30.xml emd-3545.xml | 57.8 KB 57.8 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_3545.png | 46.8 KB | ||
その他 | emd_3545_half_map_1.map.gz emd_3545_half_map_2.map.gz | 139.4 MB 139.4 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-3545 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-3545 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_3545_validation.pdf.gz | 421.4 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_3545_full_validation.pdf.gz | 420.5 KB | 表示 | |
XML形式データ | emd_3545_validation.xml.gz | 12.9 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-3545 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-3545 | HTTPS FTP |
-関連構造データ
-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_3545.map.gz / 形式: CCP4 / 大きさ: 178 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | None | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.59 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
-ハーフマップ: #1
ファイル | emd_3545_half_map_1.map | ||||||||||||
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投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: #2
ファイル | emd_3545_half_map_2.map | ||||||||||||
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投影像・断面図 |
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密度ヒストグラム |
-試料の構成要素
+全体 : Human C* Spliceosome, masked refinement
+超分子 #1: Human C* Spliceosome, masked refinement
+分子 #1: Pre-mRNA-processing-splicing factor 8
+分子 #2: 116 kDa U5 small nuclear ribonucleoprotein component
+分子 #3: SNW domain-containing protein 1
+分子 #4: Pleiotropic regulator 1
+分子 #5: Pre-mRNA-processing factor 17
+分子 #6: U5 small nuclear ribonucleoprotein 40 kDa protein
+分子 #7: Pre-mRNA-processing factor 19
+分子 #8: Pre-mRNA-splicing factor SPF27
+分子 #9: Cell division cycle 5-like protein
+分子 #10: Pre-mRNA-splicing factor SYF1
+分子 #11: Pre-mRNA-splicing factor SYF2
+分子 #12: Crooked neck-like protein 1
+分子 #13: Pre-mRNA-splicing factor RBM22
+分子 #14: Protein BUD31 homolog
+分子 #15: Spliceosome-associated protein CWC15 homolog
+分子 #16: Serine/arginine repetitive matrix protein 2
+分子 #17: Pre-mRNA-splicing factor CWC22 homolog
+分子 #18: Intron-binding protein aquarius
+分子 #19: Peptidyl-prolyl cis-trans isomerase-like 1
+分子 #20: U2 small nuclear ribonucleoprotein A'
+分子 #21: U2 small nuclear ribonucleoprotein B''
+分子 #22: Small nuclear ribonucleoprotein Sm D2
+分子 #23: Small nuclear ribonucleoprotein F
+分子 #24: Small nuclear ribonucleoprotein E
+分子 #25: Small nuclear ribonucleoprotein G
+分子 #26: Small nuclear ribonucleoprotein Sm D3
+分子 #27: Small nuclear ribonucleoprotein-associated proteins B and B'
+分子 #28: Small nuclear ribonucleoprotein Sm D1
+分子 #29: Peptidyl-prolyl cis-trans isomerase E
+分子 #30: Eukaryotic initiation factor 4A-III
+分子 #31: ATP-dependent RNA helicase DHX8
+分子 #32: MINX pre-mRNA (intron)
+分子 #33: Human gene for small nuclear RNA U2 (snRNA U2)
+分子 #34: Homo sapiens U5 A small nuclear RNA
+分子 #35: Homo sapiens RNA, U6 small nuclear 1 (RNU6-1), small nuclear RNA
-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
緩衝液 | pH: 6.4 |
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凍結 | 凍結剤: ETHANE / チャンバー内湿度: 80 % / チャンバー内温度: 277 K |
-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 II (4k x 4k) デジタル化 - 画像ごとのフレーム数: 2-17 / 平均電子線量: 2.1 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: SPOT SCAN / 撮影モード: BRIGHT FIELD / Cs: 0.01 mm |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
-画像解析
初期モデル | モデルのタイプ: OTHER / 詳細: Reconstruction from negatively stained particles |
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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: 5.9 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 136534 |
初期 角度割当 | タイプ: COMMON LINE |
最終 角度割当 | タイプ: OTHER |
-原子モデル構築 1
精密化 | 空間: REAL |
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得られたモデル | PDB-5mqf: |