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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) | |||||||||
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![]() | spliceosome / SPLICING | |||||||||
機能・相同性 | ![]() spliceosomal snRNP complex / ribonucleoprotein complex localization / snRNP binding / U4atac snRNP / positive regulation of cytotoxic T cell differentiation / maturation of 5S rRNA / RNA localization / R-loop processing / U4atac snRNA binding / box C/D sno(s)RNA binding ...spliceosomal snRNP complex / ribonucleoprotein complex localization / snRNP binding / U4atac snRNP / positive regulation of cytotoxic T cell differentiation / maturation of 5S rRNA / RNA localization / R-loop processing / U4atac snRNA binding / box C/D sno(s)RNA binding / positive regulation of primary miRNA processing / dense fibrillar component / cis assembly of pre-catalytic spliceosome / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U4/U6 snRNP / U2-type catalytic step 1 spliceosome / RNA splicing, via transesterification reactions / spliceosomal tri-snRNP complex / U4 snRNA binding / box C/D methylation guide snoRNP complex / U2-type spliceosomal complex / mRNA cis splicing, via spliceosome / U2-type precatalytic spliceosome / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / U4 snRNP / rRNA modification in the nucleus and cytosol / box C/D snoRNP assembly / U2 snRNP / U3 snoRNA binding / U2-type prespliceosome / K63-linked polyubiquitin modification-dependent protein binding / positive regulation of miRNA metabolic process / precatalytic spliceosome / spliceosomal complex assembly / mRNA Splicing - Minor Pathway / regulation of alternative mRNA splicing, via spliceosome / mRNA 3'-splice site recognition / MLL1 complex / spliceosomal tri-snRNP complex assembly / single fertilization / Major pathway of rRNA processing in the nucleolus and cytosol / U5 snRNA binding / U5 snRNP / RNA processing / U2 snRNA binding / U6 snRNA binding / spliceosomal snRNP assembly / ribonucleoprotein complex binding / pre-mRNA intronic binding / Cajal body / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / RNA splicing / mRNA Splicing - Major Pathway / helicase activity / maturation of LSU-rRNA / response to cocaine / maturation of SSU-rRNA / small-subunit processome / spliceosomal complex / mRNA processing / mRNA splicing, via spliceosome / osteoblast differentiation / cellular response to xenobiotic stimulus / protein-macromolecule adaptor activity / cellular response to tumor necrosis factor / ribosomal small subunit biogenesis / ATPase binding / cellular response to lipopolysaccharide / ubiquitinyl hydrolase 1 / cytosolic large ribosomal subunit / RNA helicase activity / hydrolase activity / RNA helicase / nuclear speck / cell cycle / cell division / intracellular membrane-bounded organelle / GTPase activity / mRNA binding / chromatin / nucleolus / GTP binding / Golgi apparatus / ATP hydrolysis activity / positive regulation of transcription by RNA polymerase II / protein-containing complex / RNA binding / zinc ion binding / extracellular exosome / nucleoplasm / ATP binding / membrane / identical protein binding / nucleus / cytoplasm / cytosol 類似検索 - 分子機能 | |||||||||
生物種 | ![]() | |||||||||
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 | |||||||||
![]() | Zhang Z / Kumar V / Dybkov O / Will CL / Zhong J / Ludwig S / Urlaub H / Kastner B / Stark H / Luehrmann R | |||||||||
資金援助 | ![]()
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![]() | ![]() 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. | |||||||||
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 | ![]() | 387.5 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 38 KB 38 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 17 KB | 表示 | ![]() |
画像 | ![]() | 48 KB | ||
Filedesc metadata | ![]() | 13.7 KB | ||
その他 | ![]() ![]() | 336.3 MB 336.3 MB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 988.5 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 988.1 KB | 表示 | |
XML形式データ | ![]() | 24.5 KB | 表示 | |
CIF形式データ | ![]() | 32.7 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | 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 ![]() |
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リンク
EMDBのページ | ![]() ![]() |
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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 | ![]() | ||||||||||||||||||||
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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:
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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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![]() | 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: ![]() |
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 |