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PLP1 alternative splicing in differentiating oligodendrocytes: Characterization of an exonic splicing enhancer

✍ Scribed by Erming Wang; Zhong Huang; Grace M. Hobson; Neviana Dimova; Karen Sperle; Andrew McCullough; Franca Cambi


Publisher
John Wiley and Sons
Year
2006
Tongue
English
Weight
541 KB
Volume
97
Category
Article
ISSN
0730-2312

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✦ Synopsis


Abstract

Proteolipid protein (PLP) and DM20 are generated by alternative splicing of exon 3B of PLP1 transcript in differentiating oligodendrocytes. We investigated the role of exonic splicing enhancers (ESE) in the selection of PLP 5β€² donor site, focusing on putative ASF/SF2, and SC35 binding motifs in exon 3B on the basis of mutations that cause disease in humans. Mutations in a putative ASF/SF2 binding motif (nucleotides 406–412) reduced PLP 5β€² donor site selection, whereas a mutation in a putative SC35 binding motif (nucleotides 382–389) had no effect. UV crosslinking and immunoprecipitation (IP) assays using an antibody to ASF/SF2 showed that the ASF/SF2 protein specifically binds to the ESE (nucleotides 406–412). The single nucleotide mutations that reduced PLP splice site selection greatly diminished ASF/SF2 protein binding to this motif. We next tested the effect of overexpressed ASF/SF2 on PLP 5β€²splice selection in differentiating oligodendrocytes. ASF/SF2 positively regulates PLP splice site selection in a concentration‐dependent manner. Disruption of the putative ASF/SF2 binding site in exon 3B reduced the positive effect of ASF/SF2 on PLP splicing. We conclude that an ESE in exon3B regulates PLP 5β€² donor site selection and that ASF/SF2 protein participates in the regulation of PLP alternative splicing in oligodendrocytes. J. Cell. Biochem. 97: 999–1016, 2006. Β© 2005 Wiley‐Liss, Inc.


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