Nucleic Acids Research, 2003, Vol. 31, No. 22 6502-6508
© 2003 Oxford University Press
Article |
Arginine/serine repeats are sufficient to constitute a splicing activation domain
Department of Genetics and Developmental Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030-3301, USA and 1 Department of Molecular and Cellular Biology, Harvard University, 7 Divinity Avenue, Cambridge, MA 02138, USA
*To whom correspondence should be addressed. Tel: +1 860 679 2090; Fax: +1 860 679 8345; Email: graveley{at}neuron.uchc.edu
Present addresses:
Alicia M. Celotto, Department of Genetics, University of Wisconsin, 445 Henry Mall, Madison, WI 53706, USA Rosa S. Tarng, Aptis Partners, 53 Burlington Road, Bedford, MA 01730, USA
SR proteins are essential pre-mRNA splicing factors that have been shown to bind a number of exonic splicing enhancers where they function to stimulate the splicing of adjacent introns. Members of the SR protein family contain one or two N-terminal RNA binding domains, as well as a C-terminal arginine–serine (RS) rich domain. The RS domains mediate protein–protein interactions with other RS domain containing proteins and are essential for many, but not all, SR protein functions. Hybrid proteins containing an RS domain fused to the bacteriophage MS2 coat protein are sufficient to activate enhancer-dependent splicing in HeLa cell nuclear extract when bound to the pre-mRNA. Here we report progress towards determining the protein sequence requirements for RS domain function. We show that the RS domains from non-SR proteins can also function as splicing activation domains when tethered to the pre-mRNA. Truncation experiments with the RS domain of the human SR protein 9G8 identified a 29 amino acid segment, containing 26 arginine or serine residues, that is sufficient to activate splicing when fused to MS2. We also show that synthetic domains composed solely of RS dipeptides are capable of activating splicing, although their potency is proportional to their size.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  V. M. Howell, J. M. Jones, S. K. Bergren, L. Li, A. C. Billi, M. R. Avenarius, and M. H. Meisler Evidence for a direct role of the disease modifier SCNM1 in splicing Hum. Mol. Genet., October 15, 2007; 16(20): 2506 - 2516. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Moroy and F. Heyd The impact of alternative splicing in vivo: Mouse models show the way RNA, August 1, 2007; 13(8): 1155 - 1171. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. P. Ng, G. Potikyan, R. O. V. Savene, C. T. Denny, V. N. Uversky, and K. A. W. Lee Multiple aromatic side chains within a disordered structure are critical for transcription and transforming activity of EWS family oncoproteins PNAS, January 9, 2007; 104(2): 479 - 484. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J.-M. Izquierdo and J. Valcarcel A simple principle to explain the evolution of pre-mRNA splicing. Genes & Dev., July 1, 2006; 20(13): 1679 - 1684. [Full Text] [PDF]
|
|
|
|
|
|
H. Shen and M. R. Green RS domains contact splicing signalsand promote splicing by a commonmechanism in yeast through humans Genes & Dev., July 1, 2006; 20(13): 1755 - 1765. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Haynes and L. M. Iakoucheva Serine/arginine-rich splicing factors belong to a class of intrinsically disordered proteins Nucleic Acids Res., January 10, 2006; 34(1): 305 - 312. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Fushimi, N. Osumi, and T. Tsukahara NSSRs/TASRs/SRp38s function as splicing modulators via binding to pre-mRNAs Genes Cells, June 1, 2005; 10(6): 531 - 541. [Abstract] [Full Text] [PDF]
|
|