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Proc Natl Acad Sci U S A. Apr 2, 1996; 93(7): 3074–3079.
PMCID: PMC39763

Pre-mRNA splicing in plants: characterization of Ser/Arg splicing factors.

Abstract

The fact that animal introns are not spliced out in plants suggests that recognition of pre-mRNA splice sites differs between the two kingdoms. In plants, little is known about proteins required for splicing, as no plant in vitro splicing system is available. Several essential splicing factors from animals, such as SF2/ASF and SC-35, belong to a family of highly conserved proteins consisting of one or two RNA binding domain(s) (RRM) and a C-terminal Ser/Arg-rich (SR or RS) domain. These animal SR proteins are required for splice site recognition and spliceosome assembly. We have screened for similar proteins in plants by using monoclonal antibodies specific for a phosphoserine epitope of the SR proteins (mAb1O4) or for SF2/ASF. These experiments demonstrate that plants do possess SR proteins, including SF2/ASF-like proteins. Similar to the animal SR proteins, this group of proteins can be isolated by two salt precipitations. However, compared to the animal SR proteins, which are highly conserved in size and number, SR proteins from Arabidopsis, carrot, and tobacco exhibit a complex pattern of intra- and interspecific variants. These plant SR proteins are able to complement inactive HeLa cell cytoplasmic S1OO extracts that are deficient in SR proteins, yielding functional splicing extracts. In addition, plant SR proteins were active in a heterologous alternative splicing assay. Thus, these plant SR proteins are authentic plant splicing factors.

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  • Nilsen TW. RNA-RNA interactions in the spliceosome: unraveling the ties that bind. Cell. 1994 Jul 15;78(1):1–4. [PubMed]
  • Birney E, Kumar S, Krainer AR. Analysis of the RNA-recognition motif and RS and RGG domains: conservation in metazoan pre-mRNA splicing factors. Nucleic Acids Res. 1993 Dec 25;21(25):5803–5816. [PMC free article] [PubMed]
  • Roth MB, Murphy C, Gall JG. A monoclonal antibody that recognizes a phosphorylated epitope stains lampbrush chromosome loops and small granules in the amphibian germinal vesicle. J Cell Biol. 1990 Dec;111(6 Pt 1):2217–2223. [PMC free article] [PubMed]
  • Zahler AM, Lane WS, Stolk JA, Roth MB. SR proteins: a conserved family of pre-mRNA splicing factors. Genes Dev. 1992 May;6(5):837–847. [PubMed]
  • Krainer AR, Conway GC, Kozak D. Purification and characterization of pre-mRNA splicing factor SF2 from HeLa cells. Genes Dev. 1990 Jul;4(7):1158–1171. [PubMed]
  • Ge H, Manley JL. A protein factor, ASF, controls cell-specific alternative splicing of SV40 early pre-mRNA in vitro. Cell. 1990 Jul 13;62(1):25–34. [PubMed]
  • Krainer AR, Mayeda A, Kozak D, Binns G. Functional expression of cloned human splicing factor SF2: homology to RNA-binding proteins, U1 70K, and Drosophila splicing regulators. Cell. 1991 Jul 26;66(2):383–394. [PubMed]
  • Ge H, Zuo P, Manley JL. Primary structure of the human splicing factor ASF reveals similarities with Drosophila regulators. Cell. 1991 Jul 26;66(2):373–382. [PubMed]
  • Mayeda A, Krainer AR. Regulation of alternative pre-mRNA splicing by hnRNP A1 and splicing factor SF2. Cell. 1992 Jan 24;68(2):365–375. [PubMed]
  • Fu XD, Maniatis T. Isolation of a complementary DNA that encodes the mammalian splicing factor SC35. Science. 1992 Apr 24;256(5056):535–538. [PubMed]
  • Fu XD, Maniatis T. The 35-kDa mammalian splicing factor SC35 mediates specific interactions between U1 and U2 small nuclear ribonucleoprotein particles at the 3' splice site. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1725–1729. [PMC free article] [PubMed]
  • Mayeda A, Zahler AM, Krainer AR, Roth MB. Two members of a conserved family of nuclear phosphoproteins are involved in pre-mRNA splicing. Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1301–1304. [PMC free article] [PubMed]
  • Krainer AR, Conway GC, Kozak D. The essential pre-mRNA splicing factor SF2 influences 5' splice site selection by activating proximal sites. Cell. 1990 Jul 13;62(1):35–42. [PubMed]
  • Fu XD, Mayeda A, Maniatis T, Krainer AR. General splicing factors SF2 and SC35 have equivalent activities in vitro, and both affect alternative 5' and 3' splice site selection. Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11224–11228. [PMC free article] [PubMed]
  • Fu XD. Specific commitment of different pre-mRNAs to splicing by single SR proteins. Nature. 1993 Sep 2;365(6441):82–85. [PubMed]
  • Li H, Bingham PM. Arginine/serine-rich domains of the su(wa) and tra RNA processing regulators target proteins to a subnuclear compartment implicated in splicing. Cell. 1991 Oct 18;67(2):335–342. [PubMed]
  • Wu JY, Maniatis T. Specific interactions between proteins implicated in splice site selection and regulated alternative splicing. Cell. 1993 Dec 17;75(6):1061–1070. [PubMed]
  • Amrein H, Hedley ML, Maniatis T. The role of specific protein-RNA and protein-protein interactions in positive and negative control of pre-mRNA splicing by Transformer 2. Cell. 1994 Feb 25;76(4):735–746. [PubMed]
  • Kohtz JD, Jamison SF, Will CL, Zuo P, Lührmann R, Garcia-Blanco MA, Manley JL. Protein-protein interactions and 5'-splice-site recognition in mammalian mRNA precursors. Nature. 1994 Mar 10;368(6467):119–124. [PubMed]
  • Brown JW, Simpson CG, Simpson GG, Turnbull-Ross AD, Clark GP. Plant pre-mRNA splicing and splicing components. Philos Trans R Soc Lond B Biol Sci. 1993 Nov 29;342(1301):217–224. [PubMed]
  • Luehrsen KR, Taha S, Walbot V. Nuclear pre-mRNA processing in higher plants. Prog Nucleic Acid Res Mol Biol. 1994;47:149–193. [PubMed]
  • Goodall GJ, Filipowicz W. The AU-rich sequences present in the introns of plant nuclear pre-mRNAs are required for splicing. Cell. 1989 Aug 11;58(3):473–483. [PubMed]
  • van Santen VL, Spritz RA. Splicing of plant pre-mRNAs in animal systems and vice versa. Gene. 1987;56(2-3):253–265. [PubMed]
  • Hunt AG, Mogen BD, Chu NM, Chua NH. The SV40 small t intron is accurately and efficiently spliced in tobacco cells. Plant Mol Biol. 1991 Mar;16(3):375–379. [PubMed]
  • Hartmuth K, Barta A. In vitro processing of a plant pre-mRNA in a HeLa cell nuclear extract. Nucleic Acids Res. 1986 Oct 10;14(19):7513–7528. [PMC free article] [PubMed]
  • Brown JW, Feix G, Frendewey D. Accurate in vitro splicing of two pre-mRNA plant introns in a HeLa cell nuclear extract. EMBO J. 1986 Nov;5(11):2749–2758. [PMC free article] [PubMed]
  • Wiebauer K, Herrero JJ, Filipowicz W. Nuclear pre-mRNA processing in plants: distinct modes of 3'-splice-site selection in plants and animals. Mol Cell Biol. 1988 May;8(5):2042–2051. [PMC free article] [PubMed]
  • Waigmann E, Barta A. Processing of chimeric introns in dicot plants: evidence for a close cooperation between 5' and 3' splice sites. Nucleic Acids Res. 1992 Jan 11;20(1):75–81. [PMC free article] [PubMed]
  • Goodall GJ, Filipowicz W. Different effects of intron nucleotide composition and secondary structure on pre-mRNA splicing in monocot and dicot plants. EMBO J. 1991 Sep;10(9):2635–2644. [PMC free article] [PubMed]
  • Simpson CG, Brown JW. Efficient splicing of an AU-rich antisense intron sequence. Plant Mol Biol. 1993 Jan;21(2):205–211. [PubMed]
  • Lou H, McCullough AJ, Schuler MA. 3' splice site selection in dicot plant nuclei is position dependent. Mol Cell Biol. 1993 Aug;13(8):4485–4493. [PMC free article] [PubMed]
  • McCullough AJ, Lou H, Schuler MA. Factors affecting authentic 5' splice site selection in plant nuclei. Mol Cell Biol. 1993 Mar;13(3):1323–1331. [PMC free article] [PubMed]
  • Luehrsen KR, Walbot V. Addition of A- and U-rich sequence increases the splicing efficiency of a deleted form of a maize intron. Plant Mol Biol. 1994 Feb;24(3):449–463. [PubMed]
  • Mayeda A, Helfman DM, Krainer AR. Modulation of exon skipping and inclusion by heterogeneous nuclear ribonucleoprotein A1 and pre-mRNA splicing factor SF2/ASF. Mol Cell Biol. 1993 May;13(5):2993–3001. [PMC free article] [PubMed]
  • Saxena SK, Ackerman EJ. Ribozymes correctly cleave a model substrate and endogenous RNA in vivo. J Biol Chem. 1990 Oct 5;265(28):17106–17109. [PubMed]
  • Roth MB, Zahler AM, Stolk JA. A conserved family of nuclear phosphoproteins localized to sites of polymerase II transcription. J Cell Biol. 1991 Nov;115(3):587–596. [PMC free article] [PubMed]
  • Krainer AR, Maniatis T, Ruskin B, Green MR. Normal and mutant human beta-globin pre-mRNAs are faithfully and efficiently spliced in vitro. Cell. 1984 Apr;36(4):993–1005. [PubMed]
  • Reed R, Maniatis T. A role for exon sequences and splice-site proximity in splice-site selection. Cell. 1986 Aug 29;46(5):681–690. [PubMed]
  • Krainer AR, Maniatis T. Multiple factors including the small nuclear ribonucleoproteins U1 and U2 are necessary for pre-mRNA splicing in vitro. Cell. 1985 Oct;42(3):725–736. [PubMed]
  • Mayeda A, Ohshima Y. Short donor site sequences inserted within the intron of beta-globin pre-mRNA serve for splicing in vitro. Mol Cell Biol. 1988 Oct;8(10):4484–4491. [PMC free article] [PubMed]
  • Krämer A, Keller W. Preparation and fractionation of mammalian extracts active in pre-mRNA splicing. Methods Enzymol. 1990;181:3–19. [PubMed]
  • Simpson GG, Vaux P, Clark G, Waugh R, Beggs JD, Brown JW. Evolutionary conservation of the spliceosomal protein, U2B''. Nucleic Acids Res. 1991 Oct 11;19(19):5213–5217. [PMC free article] [PubMed]
  • Zahler AM, Neugebauer KM, Lane WS, Roth MB. Distinct functions of SR proteins in alternative pre-mRNA splicing. Science. 1993 Apr 9;260(5105):219–222. [PubMed]
  • Lazar G, Schaal T, Maniatis T, Goodman HM. Identification of a plant serine-arginine-rich protein similar to the mammalian splicing factor SF2/ASF. Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7672–7676. [PMC free article] [PubMed]
  • Cáceres JF, Stamm S, Helfman DM, Krainer AR. Regulation of alternative splicing in vivo by overexpression of antagonistic splicing factors. Science. 1994 Sep 16;265(5179):1706–1709. [PubMed]

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