+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-4525 | |||||||||
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タイトル | Human post-catalytic spliceosome (P complex) stalled with DHX8 K594A mutant, overall map | |||||||||
マップデータ | Human post-catalytic spliceosome (P complex) stalled with DHX8 K594A mutant, overall map | |||||||||
試料 |
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機能・相同性 | 機能・相同性情報 second spliceosomal transesterification activity / exon-exon junction subcomplex mago-y14 / negative regulation of selenocysteine incorporation / regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / spliceosomal complex disassembly / cellular response to selenite ion / selenocysteine insertion sequence binding / exon-exon junction complex / pre-mRNA 3'-splice site binding / snRNP binding ...second spliceosomal transesterification activity / exon-exon junction subcomplex mago-y14 / negative regulation of selenocysteine incorporation / regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / spliceosomal complex disassembly / cellular response to selenite ion / selenocysteine insertion sequence binding / exon-exon junction complex / pre-mRNA 3'-splice site binding / snRNP binding / granulocyte differentiation / U2 snRNP binding / post-mRNA release spliceosomal complex / U7 snRNA binding / histone pre-mRNA DCP binding / regulation of retinoic acid receptor signaling pathway / U7 snRNP / negative regulation of toll-like receptor signaling pathway / 3'-5' RNA helicase activity / regulation of translation at postsynapse, modulating synaptic transmission / histone pre-mRNA 3'end processing complex / alternative mRNA splicing, via spliceosome / renal system process / cis assembly of pre-catalytic spliceosome / generation of catalytic spliceosome for first transesterification step / negative regulation of excitatory postsynaptic potential / endonucleolytic cleavage of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / regulation of vitamin D receptor signaling pathway / Z-decay: degradation of maternal mRNAs by zygotically expressed factors / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / negative regulation of interleukin-8 production / regulation of mRNA processing / negative regulation of lipopolysaccharide-mediated signaling pathway / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / Deadenylation of mRNA / embryonic brain development / protein methylation / U12-type spliceosomal complex / methylosome / negative regulation of phosphorylation / 7-methylguanosine cap hypermethylation / pre-mRNA binding / negative regulation of interferon-beta production / nuclear retinoic acid receptor binding / U2-type catalytic step 1 spliceosome / pICln-Sm protein complex / U1 snRNP binding / RNA splicing, via transesterification reactions / Prp19 complex / poly(A) binding / positive regulation of androgen receptor activity / spliceosomal tri-snRNP complex / small nuclear ribonucleoprotein complex / M-decay: degradation of maternal mRNAs by maternally stored factors / P granule / RNA stem-loop binding / sno(s)RNA-containing ribonucleoprotein complex / ATP-dependent activity, acting on RNA / SMN-Sm protein complex / mRNA 3'-end processing / mRNA cis splicing, via spliceosome / U2-type spliceosomal complex / telomerase RNA binding / embryonic cranial skeleton morphogenesis / telomerase holoenzyme complex / positive regulation of mRNA splicing, via spliceosome / U2-type precatalytic spliceosome / regulation of mRNA splicing, via spliceosome / C2H2 zinc finger domain binding / U2-type prespliceosome assembly / commitment complex / U2-type catalytic step 2 spliceosome / U4 snRNP / positive regulation by host of viral transcription / Transport of Mature mRNA derived from an Intron-Containing Transcript / positive regulation of vitamin D receptor signaling pathway / U2 snRNP / Notch binding / nuclear vitamin D receptor binding / RNA Polymerase II Transcription Termination / Regulation of gene expression in late stage (branching morphogenesis) pancreatic bud precursor cells / U1 snRNP / RUNX3 regulates NOTCH signaling / positive regulation of alpha-beta T cell differentiation / U2-type prespliceosome / NOTCH4 Intracellular Domain Regulates Transcription / K63-linked polyubiquitin modification-dependent protein binding / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / ubiquitin-ubiquitin ligase activity / exploration behavior / WD40-repeat domain binding / NOTCH3 Intracellular Domain Regulates Transcription / positive regulation of neurogenesis / lipid biosynthetic process / precatalytic spliceosome / regulation of alternative mRNA splicing, via spliceosome / hematopoietic stem cell proliferation / nuclear androgen receptor binding / cyclosporin A binding 類似検索 - 分子機能 | |||||||||
生物種 | Homo sapiens (ヒト) / Human adenovirus 2 (ヒトアデノウイルス) / 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: 3.3 Å | |||||||||
データ登録者 | Fica SM / Oubridge C / Wilkinson ME / Newman AJ / Nagai K | |||||||||
資金援助 | 英国, 2件
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引用 | ジャーナル: Science / 年: 2019 タイトル: A human postcatalytic spliceosome structure reveals essential roles of metazoan factors for exon ligation. 著者: Sebastian M Fica / Chris Oubridge / Max E Wilkinson / Andrew J Newman / Kiyoshi Nagai / 要旨: During exon ligation, the spliceosome recognizes the 3'-splice site (3'SS) of precursor messenger RNA (pre-mRNA) through non-Watson-Crick pairing with the 5'SS and the branch adenosine, in a ...During exon ligation, the spliceosome recognizes the 3'-splice site (3'SS) of precursor messenger RNA (pre-mRNA) through non-Watson-Crick pairing with the 5'SS and the branch adenosine, in a conformation stabilized by Prp18 and Prp8. Here we present the 3.3-angstrom cryo-electron microscopy structure of a human postcatalytic spliceosome just after exon ligation. The 3'SS docks at the active site through conserved RNA interactions in the absence of Prp18. Unexpectedly, the metazoan-specific FAM32A directly bridges the 5'-exon and intron 3'SS of pre-mRNA and promotes exon ligation, as shown by functional assays. CACTIN, SDE2, and NKAP-factors implicated in alternative splicing-further stabilize the catalytic conformation of the spliceosome during exon ligation. Together these four proteins act as exon ligation factors. Our study reveals how the human spliceosome has co-opted additional proteins to modulate a conserved RNA-based mechanism for 3'SS selection and to potentially fine-tune alternative splicing at the exon ligation stage. | |||||||||
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_4525.map.gz | 243.5 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-4525-v30.xml emd-4525.xml | 71 KB 71 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_4525_fsc.xml | 14.5 KB | 表示 | FSCデータファイル |
画像 | emd_4525.png | 49.1 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-4525 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-4525 | HTTPS FTP |
-関連構造データ
関連構造データ | 6qdvMC 4526C 4527C 4528C 4529C 4530C 4532C 4533C 4534C 4535C 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 (文献) |
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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_4525.map.gz / 形式: CCP4 / 大きさ: 262.9 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Human post-catalytic spliceosome (P complex) stalled with DHX8 K594A mutant, overall map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.2 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 post-catalytic P complex spliceosome
+超分子 #1: Human post-catalytic P complex spliceosome
+超分子 #2: Human post-catalytic P complex spliceosome
+超分子 #3: ATP-dependent RNA helicase DHX8
+超分子 #4: MINX introns and exons
+分子 #1: U2 snRNA
+分子 #2: U5 snRNA
+分子 #3: U6 snRNA
+分子 #11: Ligated exons: MINX mRNA
+分子 #15: Intron lariat: MINX RNA
+分子 #4: Eukaryotic initiation factor 4A-III
+分子 #5: RNA-binding protein 8A
+分子 #6: Protein mago nashi homolog 2
+分子 #7: Pre-mRNA-processing-splicing factor 8
+分子 #8: U5 small nuclear ribonucleoprotein 200 kDa helicase
+分子 #9: 116 kDa U5 small nuclear ribonucleoprotein component
+分子 #10: PRKR-interacting protein 1
+分子 #12: Cactin
+分子 #13: Protein FAM32A
+分子 #14: Pre-mRNA-splicing factor CWC22 homolog
+分子 #16: Pleiotropic regulator 1
+分子 #17: SNW domain-containing protein 1
+分子 #18: Protein BUD31 homolog
+分子 #19: Pre-mRNA-splicing factor RBM22
+分子 #20: U5 small nuclear ribonucleoprotein 40 kDa protein
+分子 #21: Cell division cycle 5-like protein
+分子 #22: Spliceosome-associated protein CWC15 homolog
+分子 #23: Serine/arginine repetitive matrix protein 2
+分子 #24: Crooked neck-like protein 1
+分子 #25: Pre-mRNA-splicing factor SYF1
+分子 #26: Intron-binding protein aquarius
+分子 #27: ATP-dependent RNA helicase DHX8
+分子 #28: U2 small nuclear ribonucleoprotein A'
+分子 #29: U2 small nuclear ribonucleoprotein B''
+分子 #30: NF-kappa-B-activating protein
+分子 #31: Small nuclear ribonucleoprotein-associated proteins B and B'
+分子 #32: Pre-mRNA-splicing factor SLU7
+分子 #33: Small nuclear ribonucleoprotein Sm D3
+分子 #34: Small nuclear ribonucleoprotein E
+分子 #35: Small nuclear ribonucleoprotein F
+分子 #36: Small nuclear ribonucleoprotein G
+分子 #37: Small nuclear ribonucleoprotein Sm D1
+分子 #38: Peptidyl-prolyl cis-trans isomerase-like 1
+分子 #39: Small nuclear ribonucleoprotein Sm D2
+分子 #40: Pre-mRNA-processing factor 17
+分子 #41: Pre-mRNA-splicing factor SPF27
+分子 #42: Pre-mRNA-processing factor 19
+分子 #43: Pre-mRNA-splicing factor SYF2
+分子 #44: Replication stress response regulator SDE2
+分子 #45: MAGNESIUM ION
+分子 #46: POTASSIUM ION
+分子 #47: ADENOSINE-5'-TRIPHOSPHATE
+分子 #48: GUANOSINE-5'-TRIPHOSPHATE
+分子 #49: ZINC ION
+分子 #50: INOSITOL HEXAKISPHOSPHATE
-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 | 0.3 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 / 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 25s, then blotted for 2.5-3.5s and immediately plunged into liquid ethane.. |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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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: FLOOD BEAM / 撮影モード: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / 最大 デフォーカス(公称値): 3.5 µm / 最小 デフォーカス(公称値): 1.5 µm / 倍率(公称値): 135000 |
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 / 撮影したグリッド数: 2 / 実像数: 6200 / 平均露光時間: 10.0 sec. / 平均電子線量: 53.0 e/Å2 |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |