The target of the DEAH-box NTP triphosphatase Prp43 in Saccharomyces cerevisiae spliceosomes is the U2 snRNP-intron interaction

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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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Jean-Baptiste Fourmann

Department of Cellular Biochemistry, Max-Planck-Institute of Biophysical Chemistry, Göttingen, Germany

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The authors declare that no competing interests exist.
Olexandr Dybkov

Department of Cellular Biochemistry, Max Planck Institute for Biophysical Chemistry, Department of Cellular Biochemistry, Germany

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The authors declare that no competing interests exist.
Dmitry E Agafonov

Department of Cellular Biochemistry, Max-Planck-Institute of Biophysical Chemistry, Göttingen, Germany

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Marcel J Tauchert

Department of Molecular Structure Biology, Institute for Microbiology and Genetics, Georg August University of Göttingen, Göttingen, Germany

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Henning Urlaub

Bioanalytical Mass Spectrometry Group, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany

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Ralf Ficner

Molecular Structure Biology, Georg August University of Göttingen, Göttingen, Germany

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Patrizia Fabrizio

Department of Cellular Biochemistry, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany

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The authors declare that no competing interests exist.
Reinhard Lührmann

Department of Cellular Biochemistry, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany

For correspondence
Reinhard.Luehrmann@mpi-bpc.mpg.de

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The authors declare that no competing interests exist.

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