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A negative element inSMN2 exon 7 inhibits splicing in spinal muscular atrophy
Nature Geneticsvolume 34, pages460–463 (2003)Cite this article
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Abstract
Spinal muscular atrophy (SMA) is a relatively common neurodegenerative disease caused by homozygous loss of the survival motor neuron 1 (SMN1) gene1. Humans possess a linked, nearly identical gene,SMN2, which produces a functional SMN protein but at levels insufficient to compensate for loss ofSMN1 (refs.1,2). A C/T transition at position +6 in exon 7 is all that differentiates the two genes, but this is sufficient to prevent efficient exon 7 splicing inSMN2 (refs.2,3). Here we show that the C/T transition functions not to disrupt an exonic splicing enhancer (ESE) inSMN1 (ref.4), as previously suggested, but rather to create an exonic splicing silencer (ESS) inSMN2. We show that this ESS functions as a binding site for a known repressor protein, hnRNP A1, which binds toSMN2 but notSMN1 exon 7 RNA. We establish the physiological importance of these results by using small interfering RNAs to reduce hnRNP A protein levels in living cells and show that this results in efficientSMN2 exon 7 splicing. Our findings not only define a new mechanism underlying the inefficient splicing ofSMN2 exon 7 but also illustrate more generally the remarkable sensitivity and precision that characterizes control of mRNA splicing.
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Acknowledgements
We thank Y. Takebayashi for technical assistance, C.L. Lorson for advice on cloningSMN1 andSMN2 minigene constructs, R. Tacke for providing human Tra2α and Tra2β cDNAs, H.J. Okano for mouse HuD cDNA, J. Kohtz for antibodies to ASF/SF2 and I. Boluk for help preparing the manuscript. This work was supported by grants from the US National Institutes of Health and Families of SMA.
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Department of Biological Sciences, Columbia University, New York, 10027, New York, USA
Tsuyoshi Kashima & James L Manley
- Tsuyoshi Kashima
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Correspondence toJames L Manley.
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Kashima, T., Manley, J. A negative element inSMN2 exon 7 inhibits splicing in spinal muscular atrophy.Nat Genet34, 460–463 (2003). https://doi.org/10.1038/ng1207
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