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The Arabidopsis splicing factor SR1 is regulated by alternative splicing

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Abstract

The serine-arginine (SR)-rich splicing factors play essential roles in general splicing and regulate alternative splice site utilization in a concentration-dependent manner. SR1 is a plant homologue of the human general/alternative splicing factor SF2/ASF. We report here that alternative splicing regulates SR1 itself. Of the five detected SR1 transcripts only one encodes the full-length protein, while the other four are different variants of the essential arginine-serine-rich domain. The data suggest that SR1 pre-mRNA could be committed to two alternate splicing pathways. One, dependent on the alternative utilization of competing 3′ splice sites in intron 9, generates SR1, SR1B and SR1C. The other, regulated by suppression of intron 9 5′ splice site utilization, generates SR1D and SR1E. The splicing pattern and molecular structure of SR1D indicates an evolutionary conservation of splicing-based regulation between plants and vertebrates and suggests that the various isoforms perform important functions. Results from transient gene expression assays indicate that alternative splicing is not an autoregulatory mechanism used to control the transcript level of the full-length protein. The ratio of SR1/SR1B transcripts, which are generated by alternative 3′ splice site utilization in intron 9, is under temperature control. The temperature-dependent increase in SR1B/SR1 ratio suggests a role of SR1B in the adaptation to high-temperature environments. In addition, based on the regulated co-expression of SR1 transcripts, it is possible that some SR1 functions could be determined by the combinatorial action of the various isoforms.

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References

  • Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Struhl, K. 1990. Current Protocols in Molecular Biology, Green & Wiley Interscience, New York.

    Google Scholar 

  • Bournay, A.S., Hedley, P.E., Maddison, A., Waugh, R. and Machray, G.C. 1996. Exon skipping induced by cold stress in a potato invertase gene transcript. Nucl. Acid Res. 24: 2347–2351.

    Google Scholar 

  • Brown, J.W.S. 1996. Arabidopsis intron mutations and pre-mRNA splicing. Plant J. 10: 771–780.

    Google Scholar 

  • Caceres, J.F. and Krainer, A.R. 1993. Functional analysis of premRNA splicing factor SF2/ASF structural domains. EMBO J. 12: 4715–4726.

    Google Scholar 

  • Church, G.M. and Kieffer-Higgins, S. 1988. Multiplex DNA sequencing. Science 240: 185–188.

    Google Scholar 

  • Fu, X.D. 1995. The superfamily of arginine/serine rich splicing factors. RNA, 1: 663–680.

    Google Scholar 

  • Gallie, D.R. and Bailey-Serres, J. 1997. Eyes off transcription! The wonderful world of post-transcriptional regulation. Plant Cell 9: 667–673.

    Google Scholar 

  • Ge, H., Zuo, P. and Manley, J.L. 1991. Primary structure of the human splicing factor ASF reveals similarities with Drosophila regulators. Cell 66: 373–382.

    Google Scholar 

  • Golovkin, M. and Reddy, A.S.N. 1996. Structure and expression of a plant U1snRNP 70K gene: alternative splicing of U1 snRNP 70K pre-mRNAs produces two different transcripts. Plant Cell 8: 1421–1435.

    Google Scholar 

  • Grabowski, P.J. 1998. Splicing regulation in neurons: tinkering with cell-specific control. Cell 92: 709–712.

    Google Scholar 

  • Grotewold, E., Prasanna, A. and Peterson, T. 1991. Alternatively spliced products of the maize P gene encode proteins with homology to the DNA binding domains of myb-like transcription factors. Proc. Natl. Acad. Sci. USA 88: 4587–4591.

    Google Scholar 

  • Haber, D.A. 1997. Splicing into senescence: the curious case of p16 and p19ARF. Cell 91: 555–558.

    Google Scholar 

  • Heinrichs, V. and Baker, B.S. 1997. In vivo analysis of the functional domains of the Drosophila splicing regulator RBP1. Proc. Natl. Acad. Sci. USA 94: 115–120.

    Google Scholar 

  • Jumaa, H. and Nielsen, P.J. 1997. The splicing factor SRp20 modifies splicing of its own mRNA and ASF/SF2 antagonizes this regulation. EMBO J. 16: 5077–5085.

    Google Scholar 

  • Kim, Y.-J., Zuo, P., Manley, J.L. and Baker, S.B. 1992. A Drosophila RNA binding protein RBP1 is localized to transcriptionally active sites of chromosomes and shows a functional similarity to human splicing factor ASF/SF2. Genes Dev. 6: 2569–2579.

    Google Scholar 

  • Kohtz, J.D., Jamison, S.F., Will, C.L., Zuo, P., Luhrmann, R., Garcia-Blanco, M.A. and Manley, J.L. 1994. Protein-protein interactions and 50 splice site recognition in mammalian mRNA precursors. Nature 368: 119–124.

    Google Scholar 

  • Krainer, A.R., Mayeda, A., Kozak, D. and Binns, G. 1991. Functional expression of cloned human splicing factor SF2: Homology to RNA-binding proteins, U170K and Drosophila splicing regulators. Cell 66: 383–394.

    Google Scholar 

  • Lazar, G., Schaal, T., Maniatis, T. and Goodman, H.M. 1995. Identification of a plant serine-arginine-rich protein similar to the mammalian splicing factor SF2/ASF. Proc. Natl. Acad. Sci. USA 92: 7672–7676.

    Google Scholar 

  • Lopato, S., Waigmann, E. and Barta, A. 1996. Characterization of a novel arginine/serine-rich splicing factor in Arabidopsis. Plant Cell 8: 2255–2264.

    Google Scholar 

  • Lopato, S., Kalyna, M., Dorner, S., Kobayashi, R., Krainer, A.R. and Barta, A. 1999. atSRp30, one of two SF2/ASF-like proteins from Arabidopsis thaliana, regulates splicing of specific plant genes. Genes Dev. 13: 987–1001.

    Google Scholar 

  • Lynch, K.W. and Maniatis, T. 1995. Synergistic interactions between two distinct elements of a regulated splicing enhancer. Genes Dev. 9: 284–293.

    Google Scholar 

  • Macknight, R., Bancroft, I., Page, T., Lister, C., Schmidt, R., Love, K., Westphal, L., Murphy, G., Sherson, S., Cobett, C. and Dean, C. 1997. FCA, a gene controlling flowering time in Arabidopsis, encodes a protein containing RNA-binding domains. Cell 89: 737–745.

    Google Scholar 

  • Manley, J.L. and Tacke, R. 1996. SR proteins and splicing control. Genes Dev. 10: 1569–1579.

    Google Scholar 

  • Mayeda, A., Zahler, A.M., Krainer, A.R. and Roth, M.B. 1992. Two members of a conserved family of nuclear phosphoproteins are involved in pre-mRNA splicing. Proc. Natl. Acad. Sci. USA 89: 1301–1304.

    Google Scholar 

  • Mermoud, J.E., Cohen, P. and Lamond, A.I. 1994. Regulation of mammalian spliceosome assembly by a protein phosphorylation mechanism. EMBO J. 13: 5679–5688.

    Google Scholar 

  • Minet, M., Dufour, M.-E. and Lacroute, F. 1992. Complementation of Saccharomyces cerevisiae auxotrophic mutants by Arabidopsis thaliana cDNAs. Plant J. 2: 417–422.

    Google Scholar 

  • Roth, M.B., Zahler, A.M. and Stolk, J.A. 1991. A conserved family of nuclear phosphoprotein is localized to sites of polymerase II transcription. J. Cell. Biol. 115: 587–596.

    Google Scholar 

  • Sablowski, R.W.M. and Meyerowitz, E.M. 1998. Temperaturesensitive splicing in the floral homeotic mutant apetala3-1. Plant Cell 10: 1453–1463.

    Google Scholar 

  • Screaton, G.R., Cacertes, J.F., Mayeda, A., Bell, M.V., Plebanski, M., Jackson, D.G., Bell, J.L. and Krainer, A.R. 1995. Identifi-cation and characterization of three members of the human SR family of pre-mRNA splicing factors. EMBO J. 14: 4336–4349.

    Google Scholar 

  • Sharp, P. and Burge, B.C. 1997. Classification of introns: U2-type or U12-type. Cell 91: 875–879.

    Google Scholar 

  • Simpson, G.G. and Filipowicz, W. 1996. Splicing of precursors to mRNA in higher plants: mechanism, regulation and sub-nuclear organization of the spliceosomal machinery. Plant Mol. Biol. 32: 1–41.

    Google Scholar 

  • Staknis, D. and Reed, R. 1994. SR proteins promote the first specific recognition of pre-mRNA and are present together with the U1 small nuclear ribonucleoprotein particle in a general splicing enhancer complex. Mol. Cell. Biol. 14: 7670–7682.

    Google Scholar 

  • Staley, J.P. and Guthrie, C. 1998. Mechanical devices of the spliceosome: motors, clocks, springs and things. Cell 92: 315–326.

    Google Scholar 

  • Sun, Q., Mayeda, A., Hampson, R.K., Krainer, A.R. and Rottman, F.M. 1993. General splicing factor SF2/A5F promotes alternative splicing by binding to an exonic splicing enhancer. Genes Dev. 7: 2598–2608.

    Google Scholar 

  • Sureau, A. and Perbal, B. 1994. Several mRNAs with variable 30 untranslated regions an different stability encode the human PR264/SC35 splicing factor Proc. Natl. Acad. Sci. USA 91: 932–936.

    Google Scholar 

  • Tacke, R. and Manley, J.L. 1995. The human splicing factors ASF/SF2 and SC35 possess different functionally significant RNA binding specificities. EMBO J. 14: 3540–3551.

    Google Scholar 

  • Tacke, R., Boned, A. and Goridis, C. 1992. ASF alternative transcripts are highly conserved between mouse and man. Nucl. Acids Res. 20: 5482.

    Google Scholar 

  • Tacke, R., Chen, Y. and Manley, J.L. 1997. Sequence-specific RNA binding by an SR protein requires RS domain phosphorylation: creation of an SRp40-specific splicing enhancer. Proc. Natl. Acad. Sci. USA 94: 1148–1153.

    Google Scholar 

  • Wu, J.Y. and Maniatis, T. 1993. Specific interactions between proteins implicated in splice site selection and regulated alternative splicing. Cell 75: 1061–1070.

    Google Scholar 

  • Xia, S.-H. and Manley, J.L. 1997. Phosphorylation of the ASF/SF2 RS domain affects both protein-protein and protein-RNA interactions and is necessary for splicing. Genes Dev. 11: 334–344.

    Google Scholar 

  • Zhang, W.J. and Wu, J.Y. 1996. Functional properties of p54, a novel SR protein active in constitutive and alternative splicing. Mol. Cell. Biol. 16: 5400–5408.

    Google Scholar 

  • Zuo, P. and Manley, J.M. 1993. Functional domains of the human splicing factor ASF/SF2. EMBO J. 12: 47727–4737.

    Google Scholar 

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Lazar, G., Goodman, H.M. The Arabidopsis splicing factor SR1 is regulated by alternative splicing. Plant Mol Biol 42, 571–581 (2000). https://doi.org/10.1023/A:1006394207479

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