Abstract
Recognition of a functional 3′ splice site in pre-mRNA splicing requires a heterodimer of the proteins U2AF65/U2AF35. U2AF65 binds to RNA at the polypyrimidine tract1,2, whereas U2AF35 is thought to interact through its arginine/serine-rich (RS) domain with other RS-domain-containing factors bound at the 5′ splice site, assembled in splicing enhancer complexes, or associated with the U4/U6.U5 small nuclear ribonucleoprotein complex3,4,5,6,7. It is unclear, however, how such network interactions can all be established through the small RS domain in U2AF35. Here we describe the function of a U2AF35-related protein (Urp), which is the human homologue of a mouse imprinted gene. Nuclear extracts depleted of Urp are defective in splicing, but activity can be restored by addition of recombinant Urp. U2AF35 could not replace Urp in complementation, indicating that their functions do not overlap. Co-immunodepletion showed that Urp is associated with the U2AF65/U2AF35 heterodimer. Binding studies revealed that Urp specifically interacts with U2AF65 through a U2AF35-homologous region and with SR proteins (a large family of RS-domain-containing proteins) through its RS domain. Therefore, Urp and U2AF35 may independently position RS-domain-containing factors within spliceosomes.
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References
Ruskin, B., Zamore, P. D. & Green, M. R. Afactor, U2AF, is required for U2 snRNP binding and splicing complex assembly. Cell 52, 207–219 (1988).
Krämer, A. The structure and function of proteins involved in mammalian pre-mRNA splicing. Annu. Rev. Biochem. 65, 367–490 (1996).
Wu, J. & Maniatis, T. Specific interactions between proteins implicated in splice site selection and regulated alternative splicing. Cell 75, 1061–1070 (1993).
Kohtz, J. D. et al. Protein–protein interactions and 5′-splice-site recognition in mammalian mRNA precursors. Nature 368, 119–124 (1994).
Zuo, P. & Maniatis, T. The splicing factor U2AF35 mediates critical protein-protein interactions in constitutive and enhancer-dependent splicing. Genes Dev. 10, 1356–1368 (1996).
Roscigno, R. F. & Garcia-Blanco, M. A. SR proteins escort the U4/U6.U5 tri-snRNP to the spliceosome. RNA 1, 692–706 (1995).
Fetzer, S., Lauber, J., Will, C. L. & Lührmann, R. The [U4/6.U5] tri-snRNP-specific 27K protein is a novel SR protein which can be phosphorylated by the snRNP-associated protein kinase. RNA 3, 344–355 (1997).
Gui, J. -F., Lane, W. S. & Fu, X. -D. Aserine kinase regulates intracellular localization of splicing factors in the cell cycle. Nature 369, 678–682 (1994).
Gui, J. -F., Tronchère, H., Chandler, S. D. & Fu, X. D. Purification and characterization of a kinase specific for the serine- and arginine-rich pre-mRNA splicing factors. Proc. Natl Acad. Sci. USA 91, 10824–10828 (1994).
Colwill, K. et al. SRPK1 and Clk/Sty protein kinases show distinct substrate specificities for serine/arginine-rich splicing factors. J. Biol. Chem. 271, 24569–24575 (1996).
Fu, X. -D. The superfamily of arginine/serine-rich splicing factors. RNA 1, 663–680 (1995).
Manley, J. L. & Tacke, R. SR proteins and splicing control. Genes Dev. 10, 1569–1579 (1996).
Kitagawa, K. et al. Isolation and mapping of human homologues of an imprinted mouse gene U2af1-rs1. Genomics 30, 257–263 (1995).
Nabetani, A., Hatada, I., Morisaki, H., Oshimura, M. & Mukai, T. Mouse U2af1-rs1 is a neomorphic imprinted gene. Mol. Cell. Biol. 17, 789–798 (1997).
Hyashizaki, Y. et al. Identification of an imprinted U2af binding protein related sequence on mouse chromosome 11 using the RLGS method. Nature Genet. 6, 33–40 (1994).
Yamaoka, T., Hatada, I., Kitagawa, K., Wang, X. & Mukai, T. Cloning and mapping of the U2af1-rs2 gene with a high transmission distortion in interspecific backcross progeny. Genomics 27, 337–340 (1995).
Zamore, P. D. & Green, M. R. Biochemical characterization of U2 snRNP auxiliary factor: an essential pre-mRNA splicing factor with novel intranuclear distribution. EMBO J. 10, 207–214 (1991).
Rudner, D. Z., Kanaar, R., Breger, K. S. & Rio, D. C. Mutations in the small subunit of the Drosphila U2AF splicing factor cause lethality and developmental defects. Proc. Natl Acad. Sci. USA 93, 10333–10337 (1996).
Zamore, P. D. & Green, M. R. Identification, purification, and biochemical characterization of U2 small nuclear ribonucleoprotein auxiliary factor. Proc. Natl Acad. Sci. USA 86, 9243–9247 (1989).
Zhang, M., Zamore, P. D., Carmo-Fonseca, M., Lamond, A. I. & Green, M. R. Cloning and intracellular localization of the U2 small nuclear ribonucleoprotein auxiliary factor small subunit. Proc. Natl Acad. Sci. USA 89, 8769–8773 (1992).
Hannon, G. J., Dmetrick, D. & Beach, D. Isolation of the Rb-related p130 through its interaction with CDK2 and cyclins. Genes Dev. 7, 2378–2391 (1993).
Harlow, E. & Lane, D. Antibodies: A Laboratory Manual(Cold Spring Harbor Laboratory Press, New York, (1988).
Zervos, A. S., Gyuris, J. & Brent, R. Mxi, a protein that specifically interacts with Max to bind Myc–Max recognition sites. Cell 72, 223–232 (1993).
Acknowledgements
We thank G. Hannon for yeast two-hybrid libraries; J.-L. Guan for raising anti-Urp antibodies; J. Fleckner and M. Green for U2AF cDNAs; P. Zuo and T. Maniatis for anti-U2AF antibodies and purified U2AF35 protein; J. Wu for two-hybrid plasmids and for advice; and members of our laboratory, especially H.-Y. Wang and J. Dyck, for suggestions and contributions. X.-D.F. is a Searle Scholar. H.T. was supported by postdoctoral fellowships from the Human Frontier Science Program and La Societé de Secours des Amis de la Science. J.W. was supported by a postdoctoral fellowship from the NIH. This work was supported by grants to X.-D.F. from the NIH.
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Tronchre, H., Wang, J. & Fu, XD. A protein related to splicing factor U2AF35 that interacts with U2AF65 and SR proteins in splicing of pre-mRNA. Nature 388, 397–400 (1997). https://doi.org/10.1038/41137
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DOI: https://doi.org/10.1038/41137
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