An erythroid differentiation–specific splicing switch in protein 4.1R mediated by the interaction of SF2/ASF with an exonic splicing enhancer
Open Access
- 1 March 2005
- journal article
- Published by American Society of Hematology in Blood
- Vol. 105 (5) , 2146-2153
- https://doi.org/10.1182/blood-2004-05-1757
Abstract
Protein 4.1R is a vital component of the red blood cell membrane cytoskeleton. Promotion of cytoskeletal junctional complex stability requires an erythroid differentiation stage–specific splicing switch promoting inclusion of exon 16 within the spectrin/actin binding domain. We showed earlier that an intricate combination of positive and negative RNA elements controls exon 16 splicing. In this report, we further identified 3 putative exonic splicing enhancers within exon 16 and investigated the function of the sequence CAGACAT in the regulation of exon 16 splicing. Mutation of these sequences leads to increased exclusion of exon 16 in both in vivo and in vitro splicing assays, indicating that CAGACAT is a functional exonic splicing enhancer. UV cross-linking further detects an approximately 33-kDa protein that specifically binds to the CAGACAT-containing transcript. An anti-SF2/ASF antibody specifically immunoprecipitates the approximately 33-kDa protein. Furthermore, SF2/ASF stimulates exon 16 inclusion in both in vitro complementation assays and minigene-transfected mouse erythroleukemia cells (MELCs). Finally, SF2/ASF expression is up-regulated and correlates with exon 16 inclusion in differentiated MELCs. These results suggest that increased splicing factor 2/alternative splicing factor (SF2/ASF) expression in differentiated mouse erythroleukemia mediates a differentiation stage–specific exon 16 splicing switch through its interaction with the exonic splicing enhancer.Keywords
This publication has 44 references indexed in Scilit:
- Alternative 5′ exons and differential splicing regulate expression of protein 4.1R isoforms with distinct N-terminiBlood, 2003
- The hnRNP A1 protein regulates HIV-1 tat splicing via a novel intron silencer elementThe EMBO Journal, 2001
- Differential Regulation of Exonic Regulatory Elements for Muscle-specific Alternative Splicing during Myogenesis and CardiogenesisPublished by Elsevier ,2000
- Binding of hnRNP H to an exonic splicing silencer is involved in the regulation of alternative splicing of the rat beta -tropomyosin geneGenes & Development, 1999
- Asynchronous regulation of splicing events within protein 4.1 pre-mRNA during erythroid differentiationBlood, 1996
- Tissue-specific alternative splicing of protein 4.1 inserts an exon necessary for formation of the ternary complex with erythrocyte spectrin and F-actinBlood, 1993
- Differentiation-associated switches in protein 4.1 expression. Synthesis of multiple structural isoforms during normal human erythropoiesis.Journal of Clinical Investigation, 1993
- Phosphorylation of protein 4.1 on tyrosine-418 modulates its function in vitro.Proceedings of the National Academy of Sciences, 1991
- Heterogeneity of mRNA and protein products arising from the protein 4.1 gene in erythroid and nonerythroid tissues.The Journal of cell biology, 1990
- Multiple protein 4.1 isoforms produced by alternative splicing in human erythroid cells.Proceedings of the National Academy of Sciences, 1988