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Three-dimensional structure of a pre-catalytic human spliceosomal complex B

Nature Structural & Molecular Biology volume 11, pages 463–468 (2004)Cite this article

Abstract

Major structural changes occur in the spliceosome during its transition from the fully assembled complex B to the catalytically activated spliceosome. To understand the rearrangement, it is necessary to know the detailed three-dimensional structures of these complexes. Here, we have immunoaffinity-purified human spliceosomes (designated BΔU1) at a stage after U4/U6·U5 tri-snRNP integration but before activation, and have determined the three-dimensional structure of BΔU1 by single-particle electron cryomicroscopy at a resolution of ∼40 Å. The overall size of the complex is about 370 × 270 × 170 Å. The three-dimensional structure features a roughly triangular body linked to a head domain in variable orientations. The body is very similar in size and shape to the isolated U4/U6·U5 tri-snRNP. This provides initial insight into the structural organization of complex B.

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Figure 1: Electron microscopy of the spliceosomal complex BΔU1.
Figure 2: Resolution of the three-dimensional reconstruction of the BΔU1 complex.
Figure 3: Determination of the absolute handedness of the BΔU1 structure by random conical tilt.
Figure 4: Surface representation of the BΔU1 complex.
Figure 5: Analysis of structural heterogeneity of the BΔU1 complex.
Figure 6: Electron microscopic visualization of the U4/U6·U5 tri-snRNP.

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Acknowledgements

We thank P. Kempkes and H. Kohansal for excellent technical assistance and the Bioverfahrenstechnik division of the Gesellschaft für Biotechnologische Forschung (GBF) in Braunschweig for large-scale HeLa cell cultivation. This work was supported by grants from the Deutsche Forschungsgemeinschaft (Lu294/12–1), the Bundesministerium für Bildung und Forschug (BMBF) (031U215B) and the Fonds der Chemischen Industrie to R.L. and a grant from BMBF (031U215B) to H.S.

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  1. Olga V Makarova

    Present address: Department of Biochemistry, University of Leicester, University Road, Leicester, LE1 7RH

Authors and Affiliations

  1. Department of Cellular Biochemistry, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, Göttingen, 37077, Germany

    Daniel Boehringer, Evgeny M Makarov, Bjoern Sander, Olga V Makarova, Berthold Kastner, Reinhard Lührmann & Holger Stark

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Correspondence to Reinhard Lührmann or Holger Stark.

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Boehringer, D., Makarov, E., Sander, B. et al. Three-dimensional structure of a pre-catalytic human spliceosomal complex B. Nat Struct Mol Biol 11, 463–468 (2004). https://doi.org/10.1038/nsmb761

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