Mutations in the proteolipid protein 1 (PLP1) gene cause the X-linked dysmyelinating diseases Pelizaeus-Merzbacher disease (PMD) and spastic paraplegia 2 (SPG2). We examined the severity of the following mutations that were suspected of affecting levels of PLP1 and DM20 RNA, the alternatively splice
PLP1 alternative splicing in differentiating oligodendrocytes: Characterization of an exonic splicing enhancer
β 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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β¦ 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 5β² 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 406β412) reduced PLP 5β² donor site selection, whereas a mutation in a putative SC35 binding motif (nucleotides 382β389) 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 406β412). 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 5β²splice selection in differentiating oligodendrocytes. ASF/SF2 positively regulates PLP splice site selection in a concentrationβdependent 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 5β² donor site selection and that ASF/SF2 protein participates in the regulation of PLP alternative splicing in oligodendrocytes. J. Cell. Biochem. 97: 999β1016, 2006. Β© 2005 WileyβLiss, Inc.
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