The target of the DEAH-box NTP triphosphatase Prp43 in Saccharomyces cerevisiae spliceosomes is the U2 snRNP-intron interaction.
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.
Department of Cellular Biochemistry, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany.
Jean-Baptiste
JB
Fourmann
Department of Cellular Biochemistry, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany.
Olexandr
O
Dybkov
Department of Cellular Biochemistry, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany.
Dmitry E
DE
Agafonov
Department of Molecular Structure Biology, Institute for Microbiology and Genetics, Georg August University of Göttingen, Göttingen, Germany.
Marcel J
MJ
Tauchert
Bionalytics, Institute for Clinical Chemistry, University Medical Center Göttingen, Göttingen, Germany.|Bioanalytical Mass Spectrometry Group, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
Henning
H
Urlaub
Department of Molecular Structure Biology, Institute for Microbiology and Genetics, Georg August University of Göttingen, Göttingen, Germany.
Ralf
R
Ficner
Department of Cellular Biochemistry, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany.
Patrizia
P
Fabrizio
Department of Cellular Biochemistry, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany.
Reinhard
0000-0002-6403-4432
R
Lührmann
D001665:Binding Sites; D053487:DEAD-box RNA Helicases / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D007438:Introns; D017413:Ribonucleoprotein, U2 Small Nuclear / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D017381:Spliceosomes / Q000378:metabolism*
C507653:Ntr2 protein, S cerevisiae; D017413:Ribonucleoprotein, U2 Small Nuclear; D029701:Saccharomyces cerevisiae Proteins; C507652:Spp382 protein, S cerevisiae; C109034:PRP43 protein, S cerevisiae; D053487:DEAD-box RNA Helicases
Human; S. cerevisiae; biochemistry; chromosomes; genes; helicases; spliceosome