Human Molecular Genetics Advance Access published online on July 6, 2005
Human Molecular Genetics, doi:10.1093/hmg/ddi233
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
1 Institute of Human Genetics, University of Newcastle upon Tyne, International Centre for Life, Central Parkway, Newcastle upon Tyne NE1 3BZ United Kingdom
* To whom correspondence should be addressed. We have discovered a new exon of the homeodomain interacting kinase HipK3 that incorporates a premature stop codon and is included only in human testis. To investigate this we tested the effects of transfecting cells with GFP fusions of RNA-binding proteins implicated in spermatogenesis using a novel assay based on multi-fraction fluorescence activated cell sorting (MF-FACS). This allows the effect of a controlled titration of any splicing factor on the splicing of endogenous genes to be studied in vivo. We found that Tra2
Received March 22, 2005
Revised June 24, 2005
Accepted June 24, 2005
Article
Upregulation of the ubiquitous alternative splicing factor Tra2
causes inclusion of a germ cell specific exon
2 Institut de Genetique et de Biologie Moleculaire et Cellulaire, CNRS/INSERM/ULP, 67404 Illkirch, C.U. Strasbourg, France
Julian P Venables, E-mail: j.venables{at}ncl.ac.uk
Abstract 
recapitulates testis-specific splicing of endogenous HipK3 in a concentration-dependent manner and binds specifically to a long purine-rich sequence in the novel exon. This sequence was also specifically bound by hnRNP A1, hnRNP H, ASF/SF2 and SRp40 but not by 9G8. Consistent with these observations in vitro studies showed this sequence shifts splicing to a downstream 5' splice site within a heterologous pre-mRNA substrate in the presence of Tra2, ASF/SF2 and SRp40, while hnRNP A1 specifically inhibits this choice. By mutating the purine-rich sequence in the context of the HipK3 gene we also show that it is the major determinant of Tra2- and hnRNP A1-mediated regulation. Tra2 is essential for sex determination and spermatogenesis in flies and Tra2 protein was most highly expressed in testis out of six mouse tissues, while hnRNP A1 is down-regulated during germ cell development. Therefore our data imply an evolutionarily conserved role for Tra2 proteins in spermatogenesis and suggest that an elevated concentration of Tra2 may convert it into a tissue-specific splicing factor.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  A. Haque, E. Buratti, and F. E. Baralle Functional properties and evolutionary splicing constraints on a composite exonic regulatory element of splicing in CFTR exon 12 Nucleic Acids Res., November 12, 2009; (2009) gkp1040v1. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Takeo, T. Kawai, K. Nishida, K. Masuda, S. Teshima-Kondo, T. Tanahashi, and K. Rokutan Oxidative stress-induced alternative splicing of transformer 2{beta} (SFRS10) and CD44 pre-mRNAs in gastric epithelial cells Am J Physiol Cell Physiol, August 1, 2009; 297(2): C330 - C338. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. J. Katzenberger, M. S. Marengo, and D. A. Wassarman Control of Alternative Splicing by Signal-dependent Degradation of Splicing-regulatory Proteins J. Biol. Chem., April 17, 2009; 284(16): 10737 - 10746. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. L. Clark, A. Coulson, C. Dalgliesh, P. Rajan, S. M. Nicol, S. Fleming, R. Heer, L. Gaughan, H. Y. Leung, D. J. Elliott, et al. The RNA Helicase p68 Is a Novel Androgen Receptor Coactivator Involved in Splicing and Is Overexpressed in Prostate Cancer Cancer Res., October 1, 2008; 68(19): 7938 - 7946. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Shukla and S. A. Fisher Tra2{beta} As a Novel Mediator of Vascular Smooth Muscle Diversification Circ. Res., August 29, 2008; 103(5): 485 - 492. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Kashima, N. Rao, C. J. David, and J. L. Manley hnRNP A1 functions with specificity in repression of SMN2 exon 7 splicing Hum. Mol. Genet., December 15, 2007; 16(24): 3149 - 3159. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. A. Sergeant, C. F. Bourgeois, C. Dalgliesh, J. P. Venables, J. Stevenin, and D. J. Elliott Alternative RNA splicing complexes containing the scaffold attachment factor SAFB2 J. Cell Sci., January 15, 2007; 120(2): 309 - 319. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Qi, S. Su, M. E. McGuffin, and W. Mattox Concentration dependent selection of targets by an SR splicing regulator results in tissue-specific RNA processing Nucleic Acids Res., December 4, 2006; 34(21): 6256 - 6263. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. J Elliott and S. N Grellscheid Alternative RNA splicing regulation in the testis. Reproduction, December 1, 2006; 132(6): 811 - 819. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Buratti, M. Baralle, and F. E. Baralle Defective splicing, disease and therapy: searching for master checkpoints in exon definition Nucleic Acids Res., July 19, 2006; 34(12): 3494 - 3510. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|