+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-32321 | |||||||||
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タイトル | The cryo-EM structure of human C* complex | |||||||||
マップデータ | The cryo-EM map of human C* complex | |||||||||
試料 |
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キーワード | spliceosome / C* complex / RNA splicing / PRP22 / exon ligation / SPLICING | |||||||||
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 / cellular response to selenite ion / spliceosomal complex disassembly / selenocysteine insertion sequence binding / exon-exon junction complex / pre-mRNA 3'-splice site binding / granulocyte differentiation ...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 / cellular response to selenite ion / spliceosomal complex disassembly / selenocysteine insertion sequence binding / exon-exon junction complex / pre-mRNA 3'-splice site binding / granulocyte differentiation / regulation of translation at postsynapse, modulating synaptic transmission / regulation of retinoic acid receptor signaling pathway / post-mRNA release spliceosomal complex / negative regulation of toll-like receptor signaling pathway / U2 snRNP binding / alternative mRNA splicing, via spliceosome / renal system process / U7 snRNA binding / negative regulation of excitatory postsynaptic potential / histone pre-mRNA DCP binding / intracellular mRNA localization / 3'-5' RNA helicase activity / negative regulation of phosphorylation / U7 snRNP / generation of catalytic spliceosome for first transesterification step / negative regulation of interleukin-8 production / Z-decay: degradation of maternal mRNAs by zygotically expressed factors / regulation of vitamin D receptor signaling pathway / histone pre-mRNA 3'end processing complex / negative regulation of lipopolysaccharide-mediated signaling pathway / cis assembly of pre-catalytic spliceosome / regulation of mRNA processing / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / Deadenylation of mRNA / embryonic cranial skeleton morphogenesis / negative regulation of interferon-beta production / protein methylation / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / 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 / positive regulation of alpha-beta T cell differentiation / 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 / negative regulation of type I interferon-mediated signaling pathway / hematopoietic stem cell proliferation / negative regulation of NF-kappaB transcription factor activity / nuclear androgen receptor binding / regulation of alternative mRNA splicing, via spliceosome / precatalytic spliceosome / K63-linked polyubiquitin modification-dependent protein binding 類似検索 - 分子機能 | |||||||||
生物種 | Homo sapiens (ヒト) / unidentified adenovirus (ウイルス) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 4.3 Å | |||||||||
データ登録者 | Zhan X / Lu Y | |||||||||
資金援助 | 中国, 1件
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引用 | ジャーナル: Mol Cell / 年: 2022 タイトル: Mechanism of exon ligation by human spliceosome. 著者: Xiechao Zhan / Yichen Lu / Xiaofeng Zhang / Chuangye Yan / Yigong Shi / 要旨: Pre-mRNA splicing involves two sequential reactions: branching and exon ligation. The C complex after branching undergoes remodeling to become the C complex, which executes exon ligation. Here, we ...Pre-mRNA splicing involves two sequential reactions: branching and exon ligation. The C complex after branching undergoes remodeling to become the C complex, which executes exon ligation. Here, we report cryo-EM structures of two intermediate human spliceosomal complexes, pre-C-I and pre-C-II, both at 3.6 Å. In both structures, the 3' splice site is already docked into the active site, the ensuing 3' exon sequences are anchored on PRP8, and the step II factor FAM192A contacts the duplex between U2 snRNA and the branch site. In the transition of pre-C-I to pre-C-II, the step II factors Cactin, FAM32A, PRKRIP1, and SLU7 are recruited. Notably, the RNA helicase PRP22 is positioned quite differently in the pre-C-I, pre-C-II, and C complexes, suggesting a role in 3' exon binding and proofreading. Together with information on human C and C complexes, our studies recapitulate a molecular choreography of the C-to-C transition, revealing mechanistic insights into exon ligation. | |||||||||
履歴 |
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-構造の表示
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 | emd_32321.map.gz | 230.3 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-32321-v30.xml emd-32321.xml | 70.4 KB 70.4 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_32321.png | 65.6 KB | ||
Filedesc metadata | emd-32321.cif.gz | 22.1 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-32321 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-32321 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_32321_validation.pdf.gz | 548.2 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_32321_full_validation.pdf.gz | 547.8 KB | 表示 | |
XML形式データ | emd_32321_validation.xml.gz | 7.3 KB | 表示 | |
CIF形式データ | emd_32321_validation.cif.gz | 8.4 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-32321 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-32321 | HTTPS FTP |
-関連構造データ
関連構造データ | 7w5bMC 7w59C 7w5aC 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 | 類似検索 - 機能・相同性F&H 検索 |
-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_32321.map.gz / 形式: CCP4 / 大きさ: 244.1 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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注釈 | The cryo-EM map of human C* complex | ||||||||||||||||||||||||||||||||||||
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: 1.338 Å | ||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
-試料の構成要素
+全体 : Human pre-C*-I complex
+超分子 #1: Human pre-C*-I complex
+分子 #1: Pre-mRNA-processing-splicing factor 8
+分子 #3: 116 kDa U5 small nuclear ribonucleoprotein component
+分子 #4: U5 small nuclear ribonucleoprotein 200 kDa helicase
+分子 #5: U5 small nuclear ribonucleoprotein 40 kDa protein
+分子 #10: Pre-mRNA-splicing factor SYF1
+分子 #11: Crooked neck-like protein 1
+分子 #12: Pre-mRNA-splicing factor SPF27
+分子 #13: Cell division cycle 5-like protein
+分子 #14: Pre-mRNA-splicing factor SYF2
+分子 #15: Protein BUD31 homolog
+分子 #16: Pre-mRNA-splicing factor RBM22
+分子 #17: Spliceosome-associated protein CWC15 homolog
+分子 #18: RNA helicase aquarius
+分子 #19: SNW domain-containing protein 1
+分子 #20: Peptidyl-prolyl cis-trans isomerase-like 1
+分子 #21: Pleiotropic regulator 1
+分子 #22: Serine/arginine repetitive matrix protein 2
+分子 #23: Pre-mRNA-splicing factor CWC22 homolog
+分子 #24: Pre-mRNA-processing factor 17
+分子 #25: ATP-dependent RNA helicase DHX8
+分子 #26: Cactin
+分子 #27: PRKR-interacting protein 1
+分子 #28: Protein FAM32A
+分子 #29: Small nuclear ribonucleoprotein-associated proteins B and B'
+分子 #30: Peptidyl-prolyl cis-trans isomerase E
+分子 #31: Small nuclear ribonucleoprotein Sm D3
+分子 #32: Small nuclear ribonucleoprotein Sm D1
+分子 #33: Small nuclear ribonucleoprotein Sm D2
+分子 #34: Small nuclear ribonucleoprotein F
+分子 #35: Small nuclear ribonucleoprotein E
+分子 #36: Small nuclear ribonucleoprotein G
+分子 #37: Pre-mRNA-processing factor 19
+分子 #38: U2 small nuclear ribonucleoprotein A'
+分子 #39: U2 small nuclear ribonucleoprotein B''
+分子 #40: Pre-mRNA-splicing factor SLU7
+分子 #41: Protein mago nashi homolog
+分子 #42: RNA-binding protein 8A
+分子 #43: Eukaryotic initiation factor 4A-III
+分子 #44: Protein CASC3
+分子 #45: NF-kappa-B-activating protein
+分子 #2: U5 snRNA
+分子 #6: U6 snRNA
+分子 #7: Pre-mRNA
+分子 #8: Pre-mRNA
+分子 #9: U2 snRNA
+分子 #46: INOSITOL HEXAKISPHOSPHATE
+分子 #47: GUANOSINE-5'-TRIPHOSPHATE
+分子 #48: MAGNESIUM ION
+分子 #49: ZINC ION
+分子 #50: ADENOSINE-5'-TRIPHOSPHATE
-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: 7.9 |
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凍結 | 凍結剤: ETHANE |
-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: GATAN K2 SUMMIT (4k x 4k) 検出モード: SUPER-RESOLUTION / 平均電子線量: 50.0 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: FLOOD BEAM / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 2.5 µm / 最小 デフォーカス(公称値): 1.2 µm |
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: INSILICO MODEL |
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最終 再構成 | 解像度のタイプ: BY AUTHOR / 解像度: 4.3 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 58374 |
初期 角度割当 | タイプ: MAXIMUM LIKELIHOOD |
最終 角度割当 | タイプ: MAXIMUM LIKELIHOOD |