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データを開く
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基本情報
| 登録情報 | ![]() | |||||||||
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| タイトル | Cryo-EM Structure of Pre-B+ATP Complex (core part) | |||||||||
マップデータ | ||||||||||
試料 |
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キーワード | spliceosome / SPLICING | |||||||||
| 機能・相同性 | 機能・相同性情報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 / box C/D sno(s)RNA binding / dense fibrillar component ...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 / box C/D sno(s)RNA binding / dense fibrillar component / cis assembly of pre-catalytic spliceosome / box C/D methylation guide snoRNP complex / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U4/U6 snRNP / RNA splicing, via transesterification reactions / U2-type catalytic step 1 spliceosome / snRNP binding / U4 snRNA binding / spliceosomal tri-snRNP complex / U2-type precatalytic spliceosome / mRNA cis splicing, via spliceosome / U2-type spliceosomal complex / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / box C/D snoRNP assembly / U4 snRNP / U2 snRNP / U3 snoRNA binding / U2-type prespliceosome / rRNA modification in the nucleus and cytosol / K63-linked polyubiquitin modification-dependent protein binding / precatalytic spliceosome / spliceosomal complex assembly / mRNA Splicing - Minor Pathway / mRNA 3'-splice site recognition / MLL1 complex / spliceosomal tri-snRNP complex assembly / U5 snRNA binding / protein deubiquitination / U5 snRNP / U2 snRNA binding / U6 snRNA binding / pre-mRNA intronic binding / single fertilization / Major pathway of rRNA processing in the nucleolus and cytosol / spliceosomal snRNP assembly / RNA processing / ribonucleoprotein complex binding / U1 snRNA binding / Cajal body / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / RNA splicing / maturation of SSU-rRNA / response to cocaine / spliceosomal complex / small-subunit processome / helicase activity / mRNA splicing, via spliceosome / cellular response to xenobiotic stimulus / mRNA processing / osteoblast differentiation / cellular response to tumor necrosis factor / cellular response to lipopolysaccharide / ATPase binding / ribosomal small subunit biogenesis / protein-macromolecule adaptor activity / ubiquitinyl hydrolase 1 / RNA helicase activity / hydrolase activity / nuclear speck / cilium / ciliary basal body / RNA helicase / cell division / intracellular membrane-bounded organelle / GTPase activity / mRNA binding / centrosome / GTP binding / chromatin / nucleolus / Golgi apparatus / positive regulation of transcription by RNA polymerase II / protein-containing complex / ATP hydrolysis activity / RNA binding / extracellular exosome / zinc ion binding / nucleoplasm / ATP binding / identical protein binding / nucleus / membrane / plasma membrane / cytosol / cytoplasm 類似検索 - 分子機能 | |||||||||
| 生物種 | Homo sapiens (ヒト) | |||||||||
| 手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.7 Å | |||||||||
データ登録者 | Zhang Z / Kumar V / Dybkov O / Will CL / Zhong J / Ludwig S / Urlaub H / Kastner B / Stark H / Luehrmann R | |||||||||
| 資金援助 | ドイツ, 1件
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引用 | ジャーナル: Nature / 年: 2024タイトル: Structural insights into the cross-exon to cross-intron spliceosome switch. 著者: Zhenwei Zhang / Vinay Kumar / Olexandr Dybkov / Cindy L Will / Jiayun Zhong / Sebastian E J Ludwig / Henning Urlaub / Berthold Kastner / Holger Stark / Reinhard Lührmann / ![]() 要旨: Early spliceosome assembly can occur through an intron-defined pathway, whereby U1 and U2 small nuclear ribonucleoprotein particles (snRNPs) assemble across the intron. Alternatively, it can occur ...Early spliceosome assembly can occur through an intron-defined pathway, whereby U1 and U2 small nuclear ribonucleoprotein particles (snRNPs) assemble across the intron. Alternatively, it can occur through an exon-defined pathway, whereby U2 binds the branch site located upstream of the defined exon and U1 snRNP interacts with the 5' splice site located directly downstream of it. The U4/U6.U5 tri-snRNP subsequently binds to produce a cross-intron (CI) or cross-exon (CE) pre-B complex, which is then converted to the spliceosomal B complex. Exon definition promotes the splicing of upstream introns and plays a key part in alternative splicing regulation. However, the three-dimensional structure of exon-defined spliceosomal complexes and the molecular mechanism of the conversion from a CE-organized to a CI-organized spliceosome, a pre-requisite for splicing catalysis, remain poorly understood. Here cryo-electron microscopy analyses of human CE pre-B complex and B-like complexes reveal extensive structural similarities with their CI counterparts. The results indicate that the CE and CI spliceosome assembly pathways converge already at the pre-B stage. Add-back experiments using purified CE pre-B complexes, coupled with cryo-electron microscopy, elucidate the order of the extensive remodelling events that accompany the formation of B complexes and B-like complexes. The molecular triggers and roles of B-specific proteins in these rearrangements are also identified. We show that CE pre-B complexes can productively bind in trans to a U1 snRNP-bound 5' splice site. Together, our studies provide new mechanistic insights into the CE to CI switch during spliceosome assembly and its effect on pre-mRNA splice site pairing at this stage. | |||||||||
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構造の表示
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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_18547.map.gz | 387.5 MB | EMDBマップデータ形式 | |
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| ヘッダ (付随情報) | emd-18547-v30.xml emd-18547.xml | 38.1 KB 38.1 KB | 表示 表示 | EMDBヘッダ |
| FSC (解像度算出) | emd_18547_fsc.xml | 17 KB | 表示 | FSCデータファイル |
| 画像 | emd_18547.png | 48 KB | ||
| Filedesc metadata | emd-18547.cif.gz | 13.7 KB | ||
| その他 | emd_18547_half_map_1.map.gz emd_18547_half_map_2.map.gz | 336.3 MB 336.3 MB | ||
| アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-18547 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-18547 | HTTPS FTP |
-検証レポート
| 文書・要旨 | emd_18547_validation.pdf.gz | 988.4 KB | 表示 | EMDB検証レポート |
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| 文書・詳細版 | emd_18547_full_validation.pdf.gz | 988 KB | 表示 | |
| XML形式データ | emd_18547_validation.xml.gz | 24.5 KB | 表示 | |
| CIF形式データ | emd_18547_validation.cif.gz | 32.7 KB | 表示 | |
| アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-18547 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-18547 | HTTPS FTP |
-関連構造データ
| 関連構造データ | ![]() 8qpbMC ![]() 8qozC ![]() 8qp8C ![]() 8qp9C ![]() 8qpaC ![]() 8qpeC ![]() 8qpkC ![]() 8qxdC ![]() 8qzsC ![]() 8r08C ![]() 8r09C ![]() 8r0aC ![]() 8r0bC ![]() 8rm5C 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 | 類似検索 - 機能・相同性 F&H 検索 |
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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 |
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マップ
| ファイル | ダウンロード / ファイル: emd_18547.map.gz / 形式: CCP4 / 大きさ: 421.9 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 | ||||||||||||||||||||||||||||||||||||
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| 対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||
| 詳細 | EMDB XML:
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-添付データ
-ハーフマップ: #2
| ファイル | emd_18547_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: #1
| ファイル | emd_18547_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 |
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試料の構成要素
+全体 : human spliceosomal pre-B+ATP complex
+超分子 #1: human spliceosomal pre-B+ATP complex
+分子 #1: U5 small nuclear ribonucleoprotein 200 kDa helicase
+分子 #2: Probable ATP-dependent RNA helicase DDX23
+分子 #3: U4/U6 small nuclear ribonucleoprotein Prp3
+分子 #4: U4/U6 small nuclear ribonucleoprotein Prp31
+分子 #5: U4/U6 small nuclear ribonucleoprotein Prp4
+分子 #6: Pre-mRNA-processing factor 6
+分子 #7: Pre-mRNA-processing-splicing factor 8
+分子 #8: Ubiquitin carboxyl-terminal hydrolase 39
+分子 #9: U4/U6.U5 tri-snRNP-associated protein 1
+分子 #10: 116 kDa U5 small nuclear ribonucleoprotein component
+分子 #11: NHP2-like protein 1, N-terminally processed
+分子 #12: Thioredoxin-like protein 4A
+分子 #14: Splicing factor 3A subunit 1
+分子 #13: U5 snRNA
+分子 #15: U4 snRNA
+分子 #16: U6 snRNA
+分子 #17: 5'ss oligo
+分子 #18: 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 |
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試料調製
| 緩衝液 | pH: 7.9 |
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| 凍結 | 凍結剤: ETHANE |
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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 III (4k x 4k) 平均電子線量: 45.0 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): 5.0 µm / 最小 デフォーカス(公称値): 1.5 µm |
| 実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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コントローラー
万見について




キーワード
Homo sapiens (ヒト)
データ登録者
ドイツ, 1件
引用




























































Z (Sec.)
Y (Row.)
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解析
FIELD EMISSION GUN

