This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Rooke, N. Articles by Black, D. L. Search for Related Content PubMed PubMed Citation Articles by Rooke, N. Articles by Black, D. L.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, March 2003, p. 1874-1884, Vol. 23, No. 6
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.6.1874-1884.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Roles for SR Proteins and hnRNP A1 in the Regulation of c-src Exon N1

Nanette Rooke,¹ Vadim Markovtsov,^1, Esra Cagavi,^1, and Douglas L. Black^1,2*

Department of Microbiology, Immunology and Molecular Genetics,¹ Howard Hughes Medical Institute, University of California, Los Angeles, Los Angeles, California 90095²

Received 16 August 2002/ Returned for modification 25 September 2002/ Accepted 16 December 2002

The splicing of the c-src exon N1 is controlled by an intricate combination of positive and negative RNA elements. Most previous work on these sequences focused on intronic elements found upstream and downstream of exon N1. However, it was demonstrated that the 5' half of the N1 exon itself acts as a splicing enhancer in vivo. Here we examine the function of this regulatory element in vitro. We show that a mutation in this sequence decreases splicing of the N1 exon in vitro. Proteins binding to this element were identified as hnRNP A1, hnRNP H, hnRNP F, and SF2/ASF by site-specific cross-linking and immunoprecipitation. The binding of these proteins to the RNA was eliminated by a mutation in the exonic element. The activities of hnRNP A1 and SF2/ASF on N1 splicing were examined by adding purified protein to in vitro splicing reactions. SF2/ASF and another SR protein, SC35, are both able to stimulate splicing of c-src pre-mRNA. However, splicing activation by SF2/ASF is dependent on the N1 exon enhancer element whereas activation by SC35 is not. In contrast to SF2/ASF and in agreement with other systems, hnRNP A1 repressed c-src splicing in vitro. The negative activity of hnRNP A1 on splicing was compared with that of PTB, a protein previously demonstrated to repress splicing in this system. Both proteins repress exon N1 splicing, and both counteract the enhancing activity of the SR proteins. Removal of the PTB binding sites upstream of N1 prevents PTB-mediated repression but does not affect A1-mediated repression. Thus, hnRNP A1 and PTB use different mechanisms to repress c-src splicing. Our results link the activity of these well-known exonic splicing regulators, SF2/ASF and hnRNP A1, to the splicing of an exon primarily controlled by intronic factors.

Present address: Rigel, Inc., South San Francisco, CA 94080.

Present address: UCLA ACCESS, Los Angeles, CA 90095.

This article has been cited by other articles:

• Mauger, D. M., Lin, C., Garcia-Blanco, M. A. (2008). hnRNP H and hnRNP F Complex with Fox2 To Silence Fibroblast Growth Factor Receptor 2 Exon IIIc. Mol. Cell. Biol. 28: 5403-5419 [Abstract] [Full Text]  
• Zhou, H.-L., Baraniak, A. P., Lou, H. (2007). Role for Fox-1/Fox-2 in Mediating the Neuronal Pathway of Calcitonin/Calcitonin Gene-Related Peptide Alternative RNA Processing. Mol. Cell. Biol. 27: 830-841 [Abstract] [Full Text]  
• Crawford, J. B., Patton, J. G. (2006). Activation of {alpha}-Tropomyosin Exon 2 Is Regulated by the SR Protein 9G8 and Heterogeneous Nuclear Ribonucleoproteins H and F. Mol. Cell. Biol. 26: 8791-8802 [Abstract] [Full Text]  
• Das, R., Dufu, K., Romney, B., Feldt, M., Elenko, M., Reed, R. (2006). Functional coupling of RNAP II transcription to spliceosome assembly.. Genes Dev. 20: 1100-1109 [Abstract] [Full Text]  
• Underwood, J. G., Boutz, P. L., Dougherty, J. D., Stoilov, P., Black, D. L. (2005). Homologues of the Caenorhabditis elegans Fox-1 Protein Are Neuronal Splicing Regulators in Mammals. Mol. Cell. Biol. 25: 10005-10016 [Abstract] [Full Text]  
• Le Sommer, C., Lesimple, M., Mereau, A., Menoret, S., Allo, M.-R., Hardy, S. (2005). PTB Regulates the Processing of a 3'-Terminal Exon by Repressing both Splicing and Polyadenylation. Mol. Cell. Biol. 25: 9595-9607 [Abstract] [Full Text]  
• Zhang, X. H.-F., Kangsamaksin, T., Chao, M. S. P., Banerjee, J. K., Chasin, L. A. (2005). Exon Inclusion Is Dependent on Predictable Exonic Splicing Enhancers. Mol. Cell. Biol. 25: 7323-7332 [Abstract] [Full Text]  
• Venables, J. P., Bourgeois, C. F., Dalgliesh, C., Kister, L., Stevenin, J., Elliott, D. J. (2005). Up-regulation of the ubiquitous alternative splicing factor Tra2{beta} causes inclusion of a germ cell-specific exon. Hum Mol Genet 14: 2289-2303 [Abstract] [Full Text]  
• Yang, G., Huang, S.-C., Wu, J. Y., Benz, E. J. Jr (2005). An erythroid differentiation-specific splicing switch in protein 4.1R mediated by the interaction of SF2/ASF with an exonic splicing enhancer. Blood 105: 2146-2153 [Abstract] [Full Text]  
• Minovitsky, S., Gee, S. L., Schokrpur, S., Dubchak, I., Conboy, J. G. (2005). The splicing regulatory element, UGCAUG, is phylogenetically and spatially conserved in introns that flank tissue-specific alternative exons. Nucleic Acids Res 33: 714-724 [Abstract] [Full Text]  
• Kralovicova, J., Houngninou-Molango, S., Kramer, A., Vorechovsky, I. (2004). Branch site haplotypes that control alternative splicing. Hum Mol Genet 13: 3189-3202 [Abstract] [Full Text]  
• Expert-Bezancon, 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. J. Biol. Chem. 279: 38249-38259 [Abstract] [Full Text]  
• Christian, K., Lang, M., Maurel, P., Raffalli-Mathieu, F. (2004). Interaction of Heterogeneous Nuclear Ribonucleoprotein A1 with Cytochrome P450 2A6 mRNA: Implications for Post-Transcriptional Regulation of the CYP2A6 Gene. Mol. Pharmacol. 65: 1405-1414 [Abstract] [Full Text]  
• Villemaire, J., Dion, I., Elela, S. A., Chabot, B. (2003). Reprogramming Alternative Pre-messenger RNA Splicing through the Use of Protein-binding Antisense Oligonucleotides. J. Biol. Chem. 278: 50031-50039 [Abstract] [Full Text]