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hnRNP A1 and the SR proteins ASF/SF2 and SC35 have antagonistic functions in splicing of beta-tropomyosin exon 6B.
J Biol Chem. 2004 Sep 10; 279(37):38249-59.JB

Abstract

Mutually exclusive splicing of exons 6A and 6B from the chicken beta-tropomyosin gene involves numerous regulatory sequences. Previously, we identified a G-rich intronic sequence (S3) downstream of exon 6B. This element consists of six G-rich motifs, mutations of which abolish splicing of exon 6B. In this paper, we investigated the cellular factors that bind to this G-rich element. By using RNA affinity chromatography, we identified heterogeneous nuclear ribonucleoprotein (hnRNP) A1, the SR proteins ASF/SF2 and SC35, and hnRNP F/H as specific components that are assembled onto the G-rich element. By using hnRNP A1-depleted HeLa nuclear extract and add-back experiments, we show that hnRNP A1 has a negative effect on splicing of exon 6B. In agreement with in vitro data, artificial recruitment of hnRNP A1, as a fusion with the MS2 coat protein, also represses splicing of exon 6B ex vivo. In contrast, ASF/SF2 and SC35 activate splicing of exon 6B. As observed with other systems, hnRNP A1 counteracts the stimulating effect of the SR proteins. Moreover, cross-linking experiments show that both ASF/SF2 and SC35 are able to displace binding of hnRNP A1 to the G-rich element, suggesting that the binding sites for these proteins are overlapping. These data indicate that the G-rich sequence is a composite element that acts as an enhancer or as a silencer, depending on which proteins bind to them.

Authors+Show Affiliations

Centre de Génétique Moléculaire, CNRS UPR 2167, Laboratoire Propre Associé à l'Université Pierre et Marie Curie, 91198 Gif-sur-Yvette, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15208309

Citation

Expert-Bezançon, Alain, et al. "HnRNP A1 and the SR Proteins ASF/SF2 and SC35 Have Antagonistic Functions in Splicing of Beta-tropomyosin Exon 6B." The Journal of Biological Chemistry, vol. 279, no. 37, 2004, pp. 38249-59.
Expert-Bezançon A, Sureau A, Durosay P, et al. HnRNP A1 and the SR proteins ASF/SF2 and SC35 have antagonistic functions in splicing of beta-tropomyosin exon 6B. J Biol Chem. 2004;279(37):38249-59.
Expert-Bezançon, A., Sureau, A., Durosay, P., Salesse, R., Groeneveld, H., Lecaer, J. P., & Marie, J. (2004). HnRNP A1 and the SR proteins ASF/SF2 and SC35 have antagonistic functions in splicing of beta-tropomyosin exon 6B. The Journal of Biological Chemistry, 279(37), 38249-59.
Expert-Bezançon A, et al. HnRNP A1 and the SR Proteins ASF/SF2 and SC35 Have Antagonistic Functions in Splicing of Beta-tropomyosin Exon 6B. J Biol Chem. 2004 Sep 10;279(37):38249-59. PubMed PMID: 15208309.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - hnRNP A1 and the SR proteins ASF/SF2 and SC35 have antagonistic functions in splicing of beta-tropomyosin exon 6B. AU - Expert-Bezançon,Alain, AU - Sureau,Alain, AU - Durosay,Patrice, AU - Salesse,Roland, AU - Groeneveld,Herman, AU - Lecaer,Jean Pierre, AU - Marie,Joëlle, Y1 - 2004/06/18/ PY - 2004/6/23/pubmed PY - 2004/10/20/medline PY - 2004/6/23/entrez SP - 38249 EP - 59 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 279 IS - 37 N2 - Mutually exclusive splicing of exons 6A and 6B from the chicken beta-tropomyosin gene involves numerous regulatory sequences. Previously, we identified a G-rich intronic sequence (S3) downstream of exon 6B. This element consists of six G-rich motifs, mutations of which abolish splicing of exon 6B. In this paper, we investigated the cellular factors that bind to this G-rich element. By using RNA affinity chromatography, we identified heterogeneous nuclear ribonucleoprotein (hnRNP) A1, the SR proteins ASF/SF2 and SC35, and hnRNP F/H as specific components that are assembled onto the G-rich element. By using hnRNP A1-depleted HeLa nuclear extract and add-back experiments, we show that hnRNP A1 has a negative effect on splicing of exon 6B. In agreement with in vitro data, artificial recruitment of hnRNP A1, as a fusion with the MS2 coat protein, also represses splicing of exon 6B ex vivo. In contrast, ASF/SF2 and SC35 activate splicing of exon 6B. As observed with other systems, hnRNP A1 counteracts the stimulating effect of the SR proteins. Moreover, cross-linking experiments show that both ASF/SF2 and SC35 are able to displace binding of hnRNP A1 to the G-rich element, suggesting that the binding sites for these proteins are overlapping. These data indicate that the G-rich sequence is a composite element that acts as an enhancer or as a silencer, depending on which proteins bind to them. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/15208309/hnRNP_A1_and_the_SR_proteins_ASF/SF2_and_SC35_have_antagonistic_functions_in_splicing_of_beta_tropomyosin_exon_6B_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=15208309 DB - PRIME DP - Unbound Medicine ER -