Published online 18 February 2004
Nucleic Acids Research, 2004, Vol. 32, No. 3 1232-1241
© 2004 Oxford University Press
ETR-3 and CELF4 protein domains required for RNA binding and splicing activity in vivo
1 Department of Pathology and 2 Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA
*To whom correspondence should be addressed. Tel: +1 713 798 3141; Fax: +1 713 798 5838; Email: tcooper{at}bcm.tmc.edu
Present address:
Nicolas Charlet-B., Institut de Genetique et de Biologie Moleculaire et Cellulaire, CNRS/INSERM/ULP, BP 10142, 67404 Illkirch Cedex, CU Strasbourg, France.
Members of the CUG-BP and ETR-3 like factor (CELF) protein family bind within conserved intronic elements (called MSEs) flanking the cardiac troponin T (cTNT) alternative exon 5 and promote exon inclusion in vivo and in vitro. Here we use a comparative deletion analysis of two family members (ETR-3 and CELF4) to identify separate domains required for RNA binding and splicing activity in vivo. CELF proteins contain two adjacent RNA binding domains (RRM1 and RRM2) near the N-terminus and one RRM (RRM3) near the C-terminus, which are separated by a 160–230 residue divergent domain of unknown function. Either RRM1 or RRM2 of CELF4 are necessary and sufficient for binding MSE RNA and RRM2 plus an additional 66 amino acids of the divergent domain are as effective as full-length protein in activating MSE-dependent splicing in vivo. Non-overlapping N- and C-terminal regions of ETR-3 containing either RRM1 and RRM2 or RRM3 plus segments of the adjacent divergent domain activate MSE-dependent exon inclusion demonstrating an unusual functional redundancy of the N- and C-termini of the protein. These results identify specific regions of ETR-3 and CELF4 that are likely targets of protein–protein interactions required for splicing activation.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  G. Natarajan, S. Ramalingam, I. Ramachandran, R. May, L. Queimado, C. W. Houchen, and S. Anant CUGBP2 downregulation by prostaglandin E2 protects colon cancer cells from radiation-induced mitotic catastrophe Am J Physiol Gastrointest Liver Physiol, May 1, 2008; 294(5): G1235 - G1244. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Subramaniam, G. Natarajan, S. Ramalingam, I. Ramachandran, R. May, L. Queimado, C. W. Houchen, and S. Anant Translation inhibition during cell cycle arrest and apoptosis: Mcl-1 is a novel target for RNA binding protein CUGBP2 Am J Physiol Gastrointest Liver Physiol, April 1, 2008; 294(4): G1025 - G1032. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. P. Chapple, K. Anthony, T. R. Martin, A. Dev, T. A. Cooper, and J.-M. Gallo Expression, localization and tau exon 10 splicing activity of the brain RNA-binding protein TNRC4 Hum. Mol. Genet., November 15, 2007; 16(22): 2760 - 2769. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Xu, C. M. Kitchen, H.-K. G. Shu, and T. J. Murphy Platelet-derived Growth Factor-induced Stabilization of Cyclooxygenase 2 mRNA in Rat Smooth Muscle Cells Requires the c-Src Family of Protein-tyrosine Kinases J. Biol. Chem., November 9, 2007; 282(45): 32699 - 32709. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. N. Ladd, G. Taffet, C. Hartley, D. L. Kearney, and T. A. Cooper Cardiac Tissue-Specific Repression of CELF Activity Disrupts Alternative Splicing and Causes Cardiomyopathy Mol. Cell. Biol., July 15, 2005; 25(14): 6267 - 6278. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Han and T. A. Cooper Identification of CELF splicing activation and repression domains in vivo Nucleic Acids Res., May 13, 2005; 33(9): 2769 - 2780. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. A. Faustino and T. A. Cooper Identification of Putative New Splicing Targets for ETR-3 Using Sequences Identified by Systematic Evolution of Ligands by Exponential Enrichment Mol. Cell. Biol., February 1, 2005; 25(3): 879 - 887. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Touchon, A. Arneodo, Y. d'Aubenton-Carafa, and C. Thermes Transcription-coupled and splicing-coupled strand asymmetries in eukaryotic genomes Nucleic Acids Res., September 23, 2004; 32(17): 4969 - 4978. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. N. Ladd and T. A. Cooper Multiple domains control the subcellular localization and activity of ETR-3, a regulator of nuclear and cytoplasmic RNA processing events J. Cell Sci., July 15, 2004; 117(16): 3519 - 3529. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. N. Ladd, N. H. Nguyen, K. Malhotra, and T. A. Cooper CELF6, a Member of the CELF Family of RNA-binding Proteins, Regulates Muscle-specific Splicing Enhancer-dependent Alternative Splicing J. Biol. Chem., April 23, 2004; 279(17): 17756 - 17764. [Abstract] [Full Text] [PDF]
|
|