A novel set of spliceosome-associated proteins and the essential splicing factor PSF bind stably to pre-mRNA prior to catalytic step II of the splicing reaction - PubMed (original) (raw)
A novel set of spliceosome-associated proteins and the essential splicing factor PSF bind stably to pre-mRNA prior to catalytic step II of the splicing reaction
O Gozani et al. EMBO J. 1994.
Abstract
We have isolated and determined the protein composition of the spliceosomal complex C. The pre-mRNA in this complex has undergone catalytic step I, but not step II, of the splicing reaction. We show that a novel set of 14 spliceosome-associated proteins (SAPs) and the essential splicing factor PSF are specifically associated with the C complex, implicating these proteins in catalytic step II. Significantly, immunodepletion and biochemical complementation studies demonstrate directly that PSF is essential for catalytic step II. Purified PSF is known to UV crosslink to pyrimidine tracts, and our data show that PSF UV crosslinks to pre-mRNA in purified C complex. Thus, PSF may replace the 3' splice site binding factor U2AF65 which is destabilized during spliceosome assembly. Finally, we show that SAPs 60 and 90, which are present in both the B and C complexes, are specifically associated with U4 and U6 snRNPs, and thus may have important roles in the functioning of these snRNPs during the splicing reaction.
Similar articles
- The splicing factor Prp17 interacts with the U2, U5 and U6 snRNPs and associates with the spliceosome pre- and post-catalysis.
Sapra AK, Khandelia P, Vijayraghavan U. Sapra AK, et al. Biochem J. 2008 Dec 15;416(3):365-74. doi: 10.1042/BJ20081195. Biochem J. 2008. PMID: 18691155 - SPF30 is an essential human splicing factor required for assembly of the U4/U5/U6 tri-small nuclear ribonucleoprotein into the spliceosome.
Rappsilber J, Ajuh P, Lamond AI, Mann M. Rappsilber J, et al. J Biol Chem. 2001 Aug 17;276(33):31142-50. doi: 10.1074/jbc.M103620200. Epub 2001 Apr 30. J Biol Chem. 2001. PMID: 11331295 - Direct interactions between pre-mRNA and six U2 small nuclear ribonucleoproteins during spliceosome assembly.
Staknis D, Reed R. Staknis D, et al. Mol Cell Biol. 1994 May;14(5):2994-3005. doi: 10.1128/mcb.14.5.2994-3005.1994. Mol Cell Biol. 1994. PMID: 8164655 Free PMC article. - Structural dynamics of the N-terminal domain and the Switch loop of Prp8 during spliceosome assembly and activation.
Jia X, Sun C. Jia X, et al. Nucleic Acids Res. 2018 May 4;46(8):3833-3840. doi: 10.1093/nar/gky242. Nucleic Acids Res. 2018. PMID: 29635373 Free PMC article. Review. - Pre-mRNA splicing: the discovery of a new spliceosome doubles the challenge.
Tarn WY, Steitz JA. Tarn WY, et al. Trends Biochem Sci. 1997 Apr;22(4):132-7. doi: 10.1016/s0968-0004(97)01018-9. Trends Biochem Sci. 1997. PMID: 9149533 Review.
Cited by
- RNA molecules display distinctive organization at nuclear speckles.
Paul S, Arias MA, Wen L, Liao SE, Zhang J, Wang X, Regev O, Fei J. Paul S, et al. iScience. 2024 Mar 27;27(5):109603. doi: 10.1016/j.isci.2024.109603. eCollection 2024 May 17. iScience. 2024. PMID: 38638569 Free PMC article. - Molecular Modelling of NONO and SFPQ Dimerization Process and RNA Recognition Mechanism.
Laurenzi T, Palazzolo L, Taiana E, Saporiti S, Ben Mariem O, Guerrini U, Neri A, Eberini I. Laurenzi T, et al. Int J Mol Sci. 2022 Jul 10;23(14):7626. doi: 10.3390/ijms23147626. Int J Mol Sci. 2022. PMID: 35886974 Free PMC article. - Multilayered control of splicing regulatory networks by DAP3 leads to widespread alternative splicing changes in cancer.
Han J, An O, Ren X, Song Y, Tang SJ, Shen H, Ke X, Ng VHE, Tay DJT, Tan HQ, Kappei D, Yang H, Chen L. Han J, et al. Nat Commun. 2022 Apr 4;13(1):1793. doi: 10.1038/s41467-022-29400-7. Nat Commun. 2022. PMID: 35379802 Free PMC article. - Epigenetic Regulation of Alternative Splicing: How LncRNAs Tailor the Message.
Pisignano G, Ladomery M. Pisignano G, et al. Noncoding RNA. 2021 Mar 11;7(1):21. doi: 10.3390/ncrna7010021. Noncoding RNA. 2021. PMID: 33799493 Free PMC article. Review. - The RNA-binding protein SFPQ preserves long-intron splicing and regulates circRNA biogenesis in mammals.
Stagsted LVW, O'Leary ET, Ebbesen KK, Hansen TB. Stagsted LVW, et al. Elife. 2021 Jan 21;10:e63088. doi: 10.7554/eLife.63088. Elife. 2021. PMID: 33476259 Free PMC article.
References
- Genes Dev. 1990 Jul;4(7):1185-96 - PubMed
- Proc Natl Acad Sci U S A. 1990 Oct;87(20):8031-5 - PubMed
- Cell. 1986 Sep 12;46(6):845-55 - PubMed
- Cell. 1986 Dec 5;47(5):755-66 - PubMed
- Cell. 1986 Dec 26;47(6):973-84 - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases