ジャーナル: Mol Cell / 年: 2010 タイトル: 3D cryo-EM structure of an active step I spliceosome and localization of its catalytic core. 著者: Monika M Golas / Bjoern Sander / Sergey Bessonov / Michael Grote / Elmar Wolf / Berthold Kastner / Holger Stark / Reinhard Lührmann / 要旨: The spliceosome excises introns from pre-mRNA in a two-step splicing reaction. So far, the three-dimensional (3D) structure of a spliceosome with preserved catalytic activity has remained elusive. ...The spliceosome excises introns from pre-mRNA in a two-step splicing reaction. So far, the three-dimensional (3D) structure of a spliceosome with preserved catalytic activity has remained elusive. Here, we determined the 3D structure of the human, catalytically active step I spliceosome (C complex) by cryo-electron microscopy (cryo-EM) in vitrified ice. Via immunolabeling we mapped the position of the 5' exon. The C complex contains an unusually salt-stable ribonucleoprotein (RNP) core that harbors its catalytic center. We determined the 3D structure of this RNP core and also that of a post-step II particle, the 35S U5 snRNP, which contains most of the C complex core proteins. As C complex domains could be recognized in these structures, their position in the C complex could be determined, thereby allowing the region harboring the spliceosome's catalytic core to be localized.
全体 : High salt resistant human spliceosomal C complex core
全体
名称: High salt resistant human spliceosomal C complex core
要素
試料: High salt resistant human spliceosomal C complex core
細胞器官・細胞要素: High salt resistant human spliceosomal C complex core
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超分子 #1000: High salt resistant human spliceosomal C complex core
超分子
名称: High salt resistant human spliceosomal C complex core タイプ: sample / ID: 1000 / Number unique components: 1
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超分子 #1: High salt resistant human spliceosomal C complex core
超分子
名称: High salt resistant human spliceosomal C complex core タイプ: organelle_or_cellular_component / ID: 1 / Name.synonym: High salt core of spliceosomal C complex / 組換発現: No
由来(天然)
生物種: Homo sapiens (ヒト) / 別称: Human
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実験情報
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構造解析
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
凍結
凍結剤: NONE / 装置: OTHER
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電子顕微鏡法
顕微鏡
FEI TECNAI 12
電子線
加速電圧: 120 kV / 電子線源: LAB6
電子光学系
照射モード: SPOT SCAN / 撮影モード: BRIGHT FIELD
試料ステージ
試料ホルダー: Eucentric / 試料ホルダーモデル: SIDE ENTRY, EUCENTRIC