Arginine methylation analysis of the splicing-associated SR protein SFRS9-SRP30C

• Autorzy: Bressan G C , Moraes E.C. , Monfiolli A.O. , Kuniyoshi T.M. , Passos D.O. , Gomes M.D. , Kobarg J.
• Języki publikacji: EN

Abstrakty

EN

The human SFRS9/SRp30c belongs to the SR family of splicing regulators. Despite evidence that members of this protein family may be targeted by arginine methylation, this has yet to be experimentally addressed. In this study, we found that SFRS9 is a target for PRMT1-mediated arginine methylation in vitro, and that it is immunoprecipitated from HEK-293 lysates by antibodies that recognize both mono- and dimethylated arginines. We further observed that upon treatment with the methylation inhibitor Adox, the fluorescent EGFP-SFRS9 re-localizes to dot-like structures in the cell nucleus. In subsequent confocal analyses, we found that EGFP-SFRS9 localizes to nucleoli in Adox-treated cells. Our findings indicate the importance of arginine methylation for the subnuclear localization of SFRS9.

Słowa kluczowe

EN

arginine methylation; protein-protein interaction; nuclear body; monoclonal antibody

• Wydawca: -
• Czasopismo: Cellular and Molecular Biology Letters
• Rocznik: 2009
• Tom: 14
• Numer: 4
• Strony: 657-669
• Opis fizyczny: p.657-669,fig.,ref.

Twórcy

autor

Bressan G C

• Universidade Estadual de Campinas, Campinas, SP, Brasil

autor

Moraes E.C.

autor

Monfiolli A.O.

autor

Kuniyoshi T.M.

autor

Passos D.O.

autor

Gomes M.D.

autor

Kobarg J.

Bibliografia

• 1. Wang, Z. and Burge, C.B. Splicing regulation: From a parts list of regulatory elements to an integrated splicing code. RNA 14 (2008) 802-813.
• 2. Reed, R. Initial splice-site recognition and pairing during pre-mRNA splicing. Curr. Opin. Genet. Dev. 6 (1996) 215-220.
• 3. Chew, S.L., Liu, H.X., Mayeda, A. and Krainer, A.R. Evidence for the function of an exonic splicing enhancer after the first catalytic step of premRNA splicing. Proc. Natl. Acad. Sci. USA 96 (1999) 10655-10660.
• 4. Hertel, K.J. and Graveley, B.R. RS domains contact the pre-mRNA throughout spliceosome assembly.Trends Biochem. Sci. 30 (2005) 115-118.
• 5. Boisvert, F.M., Côté, J., Boulanger, M.C. and Richard, S. A proteomic analysis of arginine-methylated protein complexes. Mol. Cell. Proteomics 2 (2003) 1319-1330.
• 6. Godin, K.S. and Varani, G. How arginine-rich domains coordinate mRNA maturation events. RNA Biol. 4 (2007) 69-75.
• 7. Letunic, I., Doerks, T., and Bork, P. SMART 6: recent updates and new developments. Nucleic Acids Res. 37 (2009) D229-322.
• 8. Passos, D.O., Bressan, G.C., Nery, F.C. and Kobarg, J. Ki-1/57 interacts with PRMT1 and is a substrate for arginine methylation. FEBS J. 273 (2006) 3946-3961.
• 9. Passos. D.O., Quaresma, J.C. and Kobarg, J. The methylation of the C-terminal region of hnRNPQ (NSAP1) is important for its nuclear localization. Biochem. Biophys. Res. Commun. 346 (2006) 517-525.
• 10. De Leeuw, F., Zhang, T., Wauquier, C., Huez, G., Kruys, V. and Gueydan C. The cold-inducible RNA-binding protein migrates from the nucleus to cytoplasmic stress granules by a methylation-dependent mechanism and acts as a translational repressor. Exp. Cell. Res. 313 (2007) 4130-4144.
• 11. Cheng, D., Côté, J., Shaaban, S. and Bedford, M.T. The arginine methyltransferase CARM1 regulates the coupling of transcription and mRNA processing. Mol. Cell. 25 (2007) 71-83.
• 12. Petersen-Mahrt, S.K., Estmer, C., Ohrmalm, C., Matthews, D.A., Russell, W.C. and Akusjärvi, G. The splicing factor-associated protein, p32, regulates RNA splicing by inhibiting ASF/SF2 RNA binding and phosphorylation. EMBO J. 18 (1999) 1014-1024.
• 13. Nery, F.C., Rui, E., Kuniyoshi, T.M. and Kobarg, J. Evidence for the interaction of the regulatory protein Ki-1⁄57 with p53 and its interacting proteins. Biochem. Biophys. Res. Commun. 341 (2006) 847-855.
• 14. Tadesse, H., Deschênes-Furry, J., Boisvenue, S. and Côté, J. KH-type splicing regulatory protein interacts with survival motor neuron protein and is misregulated in spinal muscular atrophy. Hum. Mol. Genet. 17 (2008) 506-524.
• 15. Raffetseder, U., Frye, B., Rauen, T., Jürchott, K., Royer, H.D., Jansen, P.L. and Mertens, P.R. Splicing factor SRp30c interaction with Y-box protein-1 confers nuclear YB-1 shuttling and alternative splice site selection. J. Biol. Chem. 278 (2003) 18241-18248.
• 16. Lyon, C.E., Bohmann, K., Sleeman, J. and Lamond, A.I. Inhibition of protein dephosphorylation results in the accumulation of splicing snRNPs and coiled bodies within the nucleolus. Exp. Cell. Res. 230 (1997) 84-93.
• 17. Lamond, A.I. and Spector, D.L. Nuclear speckles: a model for nuclear organelles. Nat. Rev. Mol. Cell. Biol. 4 (2003) 605-612.
• 18. Boisvert, F.M., van Koningsbruggen, S., Navascués, J. and Lamond, A.I. The multifunctional nucleolus. Nat. Rev. Mol. Cell. Biol. 8 (2007) 574-585.
• 19. Gerbi, S.A., Borovjagin, A.V. and Lange, T.S. The nucleolus: a site of ribonucleoprotein maturation. Curr. Opin. Cell Biol. 15 (2003) 318-325.
• 20. Lange, T.S. and Gerbi, S.A. Transient nucleolar localization of U6 small nuclear RNA. Mol. Biol. Cell 11 (2000) 2419-2428.
• 21. Gerbi, S.A. and Lange, T.S. All small nuclear RNAs (snRNAs) of the (U4/U6.U5) tri-snRNP localize to nucleoli; identification of the nucleolar localization element of U6 snRNA. Mol. Biol. Cell 13 (2002) 3123-3137.
• 22. Lai, M.C., Kuo, H.W., Chang, W.C. and Tarn, W.Y. A novel splicing regulator shares a nuclear import pathway with SR proteins. EMBO J. 22 (2003) 1359-1369.
• 23. Wagner, S., Chiosea, S. and Nickerson, J. A. The spatial targeting and nuclear matrix binding domains of SRm160. Proc. Natl. Acad. Sci. U.S.A. 100 (2003) 3269-3274.
• 24. Andersen, J.S., Lyon, C.E., Fox, A.H., Leung, A.K., Lam, Y.W., Steen, H.., Mann, M. and Lamond, A.I. Directed proteomic analysis of the human nucleolus. Curr. Biol. 12 (2002) 1-11.
• 25. Leung, A.K., Andersen, J.S., Mann, M. and Lamond, A.I. Bioinformatic analysis of the nucleolus. Biochem. J. 376 (2003) 553-569.
• 26. Larkin, M.A., Blackshields, G., Brown, N.P., Chenna, R., McGettigan, P.A., McWilliam, H., Valentin, F., Wallace, I.M., Wilm, A., Lopez, R., Thompson, J.D., Gibson, T.J. and Higgins, D.G. ClustalW2 and ClustalX version 2. Bioinformatics 23 (2007) 2947-2948.
• 27. Chen, H., Xue, Y., Huang, N., Yao, X. and Sun, Z. MeMo: a web tool for prediction of protein methylation modifications. Nucleic Acids Res. 34 (2006) 249-253.
• 28. Assmann, E.M., Alborghetti, M.R., Camargo, M.E. and Kobarg, J. FEZ1 dimerization and interaction with transcription regulatory proteins involves its coiled-coil region. J. Biol. Chem. 281 (2006) 9869-9881.