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  SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing.

Pozzi, B., Bragado, L., Will, C. L., Mammi, P., Risso, G., Urlaub, H., et al. (2017). SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing. Nucleic Acids Research, 45(11), 6729-6745. doi:10.1093/nar/gkx213.

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Pozzi, B., Author
Bragado, L., Author
Will, C. L.1, Author           
Mammi, P., Author
Risso, G., Author
Urlaub, H.2, Author           
Lührmann, R.1, Author           
Srebrow, A., Author
Affiliations:
1Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578576              
2Research Group of Bioanalytical Mass Spectrometry, MPI for biophysical chemistry, Max Planck Society, ou_578613              

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 Abstract: Pre-mRNA splicing is catalyzed by the spliceosome, a multi-megadalton ribonucleoprotein machine. Previous work from our laboratory revealed the splicing factor SRSF1 as a regulator of the SUMO pathway, leading us to explore a connection between this pathway and the splicing machinery. We show here that addition of a recombinant SUMO-protease decreases the efficiency of pre-mRNA splicing in vitro. By mass spectrometry analysis of anti-SUMO immunoprecipitated proteins obtained from purified splicing complexes formed along the splicing reaction, we identified spliceosome-associated SUMO substrates. After corroborating SUMOylation of Prp3 in cultured cells, we defined Lys 289 and Lys 559 as bona fide SUMO attachment sites within this spliceosomal protein. We further demonstrated that a Prp3 SUMOylation-deficient mutant while still capable of interacting with U4/U6 snRNP components, is unable to co-precipitate U2 and U5 snRNA and the spliceosomal proteins U2-SF3a120 and U5-Snu114. This SUMOylation-deficient mutant fails to restore the splicing of different pre-mRNAs to the levels achieved by the wild type protein, when transfected into Prp3-depleted cultured cells. This mutant also shows a diminished recruitment to active spliceosomes, compared to the wild type protein. These findings indicate that SUMO conjugation plays a role during the splicing process and suggest the involvement of Prp3 SUMOylation in U4/U6•U5 tri-snRNP formation and/or recruitment.

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Language(s): eng - English
 Dates: 2017-03-302017
 Publication Status: Issued
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 Rev. Type: Peer
 Identifiers: DOI: 10.1093/nar/gkx213
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Title: Nucleic Acids Research
Source Genre: Journal
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Pages: - Volume / Issue: 45 (11) Sequence Number: - Start / End Page: 6729 - 6745 Identifier: -