This Article Full Text Full Text (PDF) All Versions of this Article: 16/11/2426    most recent Author Manuscript (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Seong, J. Y. Articles by Kim, K. Search for Related Content PubMed PubMed Citation Articles by Seong, J. Y. Articles by Kim, K. Pubmed/NCBI databases Gene GEO Profiles HomoloGene Nucleotide Protein UniGene Compound via MeSH Substance via MeSH Hazardous Substances DB (L)-ARGININE L-SERINE

Exonic Splicing Enhancer-Dependent Splicing of the Gonadotropin-Releasing Hormone Premessenger Ribonucleic Acid Is Mediated by Tra2, a 40-Kilodalton Serine/Arginine-Rich Protein

Hormone Research Center (J.Y.S.), Chonnam National University, Kwangju 500-757, Korea; School of Biological Sciences (J.H., S.P., K.K.), Seoul National University, Seoul 151-742, Korea; and Department of Obstetrics and Gynecology (W.W., H.J.), University of Göttingen, Göttingen 37075, Germany

Address all correspondence and requests for reprints to: Kyungjin Kim, Ph.D., School of Biological Sciences Seoul National University Seoul 151-742, Korea. E-mail: .

In an earlier study, we found that excision of the first intron (intron A) from the rat GnRH primary transcript is attenuated in non-GnRH-producing cells. This attenuation can be partially relieved by exonic splicing enhancers (ESEs) located in GnRH exons 3 and 4. In the present study, we confirmed that intron A of the mouse GnRH pre-mRNA was not excised in a HeLa nuclear extract (NE) in vitro or in COS-7 cells in vivo. Intron A could, however, be partially removed when exon 3 and/or 4 were linked to exon 2. In the presence of an ESE in exon 4 (ESE4), an addition of GT1 NE further increased the excision rate of intron A, whereas the addition of KK1 (a non-GnRH-producing cell) NE decreased it. To define the GnRH neuron-specific splicing activity, GT1 NE was fractionated by ultracentrifugation and ammonium sulfate precipitation. A 50–90% ammonium sulfate pellet (ASP50–90) fraction was further precipitated with 20 mM MgCl₂ to isolate a serine/arginine-rich (SR) protein fraction. Among the ASP fractions, ASP40–50 significantly increased the excision rate of intron A in the presence of HeLa NE or SR protein-rich fraction. However, the ASP40–50 fraction alone could not remove intron A. This result suggests the presence of a cofactor protein(s) in the ASP40–50 fraction that may mediate the interaction between a 3' spliceosome complex and the ESE4-SR protein complex. UV cross-linking and gel mobility shift analysis revealed that Tra2 but not other SR proteins tested, specifically binds to ESE4. Moreover, Tra2 stimulated intron A excision in a dose-dependent manner. These results imply that Tra2 and a cofactor protein in the ASP40–50 fraction are involved in mediating the GnRH neuron-specific excision of intron A from the GnRH primary transcript.

This article has been cited by other articles:

				A. Disset, C.F. Bourgeois, N. Benmalek, M. Claustres, J. Stevenin, and S. Tuffery-Giraud An exon skipping-associated nonsense mutation in the dystrophin gene uncovers a complex interplay between multiple antagonistic splicing elements Hum. Mol. Genet., March 15, 2006; 15(6): 999 - 1013. [Abstract] [Full Text] [PDF]

				E. Park, J. Han, G. H. Son, M. S. Lee, S. Chung, S. H. Park, K. Park, K. H. Lee, S. Choi, J. Y. Seong, et al. Cooperative Actions of Tra2{alpha} with 9G8 and SRp30c in the RNA Splicing of the Gonadotropin-releasing Hormone Gene Transcript J. Biol. Chem., January 6, 2006; 281(1): 401 - 409. [Abstract] [Full Text] [PDF]

				J. P. Venables, C. F. Bourgeois, C. Dalgliesh, L. Kister, J. Stevenin, and D. J. Elliott Up-regulation of the ubiquitous alternative splicing factor Tra2{beta} causes inclusion of a germ cell-specific exon Hum. Mol. Genet., August 15, 2005; 14(16): 2289 - 2303. [Abstract] [Full Text] [PDF]

				G. H. Son, H. Jung, J. Y. Seong, Y. Choe, D. Geum, and K. Kim Excision of the First Intron from the Gonadotropin-releasing Hormone (GnRH) Transcript Serves as a Key Regulatory Step for GnRH Biosynthesis J. Biol. Chem., May 9, 2003; 278(20): 18037 - 18044. [Abstract] [Full Text] [PDF]