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Elife. 2016 Apr 26;5. pii: e15564. doi: 10.7554/eLife.15564.

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

Author information

1
Department of Cellular Biochemistry, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany.
2
Department of Molecular Structure Biology, Institute for Microbiology and Genetics, Georg August University of Göttingen, Göttingen, Germany.
3
Bionalytics, Institute for Clinical Chemistry, University Medical Center Göttingen, Göttingen, Germany.
4
Bioanalytical Mass Spectrometry Group, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.

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, B(act)), 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.

KEYWORDS:

Human; S. cerevisiae; biochemistry; chromosomes; genes; helicases; spliceosome

PMID:
27115347
PMCID:
PMC4866824
DOI:
10.7554/eLife.15564
[Indexed for MEDLINE]
Free PMC Article

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