1. Biochemistry and Chemical Biology
  2. Chromosomes and Gene Expression
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The target of the DEAH-box NTP triphosphatase Prp43 in Saccharomyces cerevisiae spliceosomes is the U2 snRNP-intron interaction

  1. Jean-Baptiste Fourmann
  2. Olexandr Dybkov
  3. Dmitry E Agafonov
  4. Marcel J Tauchert
  5. Henning Urlaub
  6. Ralf Ficner
  7. Patrizia Fabrizio
  8. Reinhard Lührmann  Is a corresponding author
  1. Max-Planck-Institute of Biophysical Chemistry, Germany
  2. Max Planck Institute for Biophysical Chemistry, Germany
  3. Georg August University of Göttingen, Germany
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Cite this article as: eLife 2016;5:e15564 doi: 10.7554/eLife.15564

Abstract

The DEAH-box NTPase Prp43 and its cofactors Ntr1 and Ntr2 form the NTR complex and are required for disassembling intron-lariat spliceosomes (ILS) and defective earlier spliceosomes. However, the Prp43 binding site in the spliceosome and its target(s) are unknown. We show that Prp43 fused to Ntr1's G patch motif (Prp43_Ntr1GP) is as efficient as the NTR in ILS disassembly, yielding identical dissociation products and recognizing its natural ILS target even in the absence of Ntr1's C-terminal-domain (CTD) and Ntr2. Unlike the NTR, Prp43_Ntr1GP disassembles earlier spliceosomal complexes (A, B, Bact), indicating that Ntr2/Ntr1-CTD prevents NTR from disrupting properly assembled spliceosomes other than the ILS. The U2 snRNP-intron interaction is disrupted in all complexes by Prp43_Ntr1GP, and in the spliceosome contacts U2 proteins and the pre-mRNA, indicating that the U2 snRNP-intron interaction is Prp43's major target.

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Author details

  1. Jean-Baptiste Fourmann

    Department of Cellular Biochemistry, Max-Planck-Institute of Biophysical Chemistry, Göttingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Olexandr Dybkov

    Department of Cellular Biochemistry, Max Planck Institute for Biophysical Chemistry, Department of Cellular Biochemistry, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Dmitry E Agafonov

    Department of Cellular Biochemistry, Max-Planck-Institute of Biophysical Chemistry, Göttingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. Marcel J Tauchert

    Department of Molecular Structure Biology, Institute for Microbiology and Genetics, Georg August University of Göttingen, Göttingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. Henning Urlaub

    Bioanalytical Mass Spectrometry Group, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Ralf Ficner

    Molecular Structure Biology, Georg August University of Göttingen, Göttingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  7. Patrizia Fabrizio

    Department of Cellular Biochemistry, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Reinhard Lührmann

    Department of Cellular Biochemistry, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany
    For correspondence
    Reinhard.Luehrmann@mpi-bpc.mpg.de
    Competing interests
    The authors declare that no competing interests exist.

Reviewing Editor

  1. Timothy W Nilsen, Case Western Reserve University, United States

Publication history

  1. Received: February 25, 2016
  2. Accepted: April 25, 2016
  3. Accepted Manuscript published: April 26, 2016 (version 1)
  4. Accepted Manuscript updated: April 27, 2016 (version 2)
  5. Version of Record published: May 13, 2016 (version 3)

Copyright

© 2016, Fourmann et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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