SRp54 (SFRS11), a regulator for tau exon 10 alternative splicing identified by an expression cloning strategy.
The tau gene encodes a microtubule-associated protein that is critical for neuronal survival and function. Splicing defects in the human tau gene lead to frontotemporal dementia with Parkinsonism linked to chromosome 17 (FTDP-17), an autosomal dominant neurodegenerative disorder. Genetic mutations associated with FTDP-17 often affect tau exon 10 alternative splicing. To investigate mechanisms regulating tau exon 10 alternative splicing, we have developed a green fluorescent protein reporter for tau exon 10 skipping and an expression cloning strategy to identify splicing regulators. A role for SRp54 (also named SFRS11) as a tau exon 10 splicing repressor has been uncovered using this strategy. The overexpression of SRp54 suppresses tau exon 10 inclusion. RNA interference-mediated knock-down of SRp54 increases exon 10 inclusion. SRp54 interacts with a purine-rich element in exon 10 and antagonizes Tra2beta, an SR-domain-containing protein that enhances exon 10 inclusion. Deletion of this exonic element eliminates the activity of SRp54 in suppressing exon 10 inclusion. Our data support a role of SRp54 in regulating tau exon 10 splicing. These experiments also establish a generally useful approach for identifying trans-acting regulators of alternative splicing by expression cloning.
Wu
Jane Y
JY
Northwestern University Feinberg School of Medicine, Center for Genetic Medicine, 303 E. Superior St., Lurie 6-117, Chicago, IL 60611, USA. jane-wu@northwestern.edu
Kar
Amar
A
Kuo
David
D
Yu
Bing
B
Havlioglu
Necat
N
D017398:Alternative Splicing / Q000235:genetics*; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods*; D015536:Down-Regulation / Q000235:genetics; D004742:Enhancer Elements, Genetic / Q000235:genetics; D005091:Exons / Q000235:genetics*; D005333:Fetus / Q000378:metabolism; D015870:Gene Expression; D015723:Gene Library; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D051916:Membrane Cofactor Protein / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011687:Purines / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion / Q000235:genetics; D000068103:Serine-Arginine Splicing Factors; D015533:Transcriptional Activation / Q000235:genetics; D016875:tau Proteins / Q000235:genetics*
C497857:CD46 protein, human; D051916:Membrane Cofactor Protein; D009687:Nuclear Proteins; D011687:Purines; D012333:RNA, Messenger; C101745:Sfrs11 protein; D016875:tau Proteins; D049452:Green Fluorescent Proteins; D000068103:Serine-Arginine Splicing Factors