Abstract
Spinal muscular atrophy (SMA) is the leading genetic cause of infant mortality. SMA is caused by the homozygous absence of survival motor neuron-1 (SMN1). SMN2, a nearly identical copy gene, is retained in all SMA patients and encodes an identical protein as SMN1; however, SMN1 and SMN2 differ by a silent C to T transition which results in the production of an alternatively spliced isoform (SMNDelta7), which encodes a defective protein, demonstrating that the absence of the short peptide encoded by SMN exon 7 is critical in SMA development. Previously, we have shown that for some functions heterologous sequences can compensate for the exon 7 peptide, suggesting that the SMN C-terminus functions non-specifically. Consistent with this hypothesis, we now identify novel aminoglycosides that can induce SMN protein levels in patient fibroblasts. This hypothesis was supported, in part, by a novel fluorescent SMN read-through assay. Interestingly, however, through the development of a SMN exon 7-specific antibody, results suggested that levels of normal full-length SMN might also be elevated by aminoglycoside treatment. These results demonstrate that the compounds that promote read-through may provide an alternative platform for the discovery of compounds that induce SMN protein levels.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Aminoglycosides / chemistry
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Aminoglycosides / pharmacology*
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Blotting, Western
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Cells, Cultured
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Cyclic AMP Response Element-Binding Protein / genetics*
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Cyclic AMP Response Element-Binding Protein / metabolism
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Dose-Response Relationship, Drug
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Enzyme Inhibitors / pharmacology
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Exons
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Fibroblasts / drug effects*
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Fibroblasts / metabolism
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Fibroblasts / pathology
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Gene Expression / drug effects
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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HeLa Cells
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Histone Acetyltransferases / antagonists & inhibitors
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Humans
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Luciferases / genetics
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Luciferases / metabolism
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Molecular Structure
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Muscular Atrophy, Spinal / genetics
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Muscular Atrophy, Spinal / metabolism
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Muscular Atrophy, Spinal / pathology
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Nerve Tissue Proteins / genetics*
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Nerve Tissue Proteins / metabolism
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RNA Splicing / drug effects
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RNA-Binding Proteins / genetics*
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RNA-Binding Proteins / metabolism
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Reverse Transcriptase Polymerase Chain Reaction
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SMN Complex Proteins
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Survival of Motor Neuron 1 Protein
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Survival of Motor Neuron 2 Protein
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Time Factors
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Transfection
Substances
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Aminoglycosides
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Cyclic AMP Response Element-Binding Protein
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Enzyme Inhibitors
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Nerve Tissue Proteins
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RNA-Binding Proteins
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Recombinant Fusion Proteins
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SMN Complex Proteins
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SMN1 protein, human
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SMN2 protein, human
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Survival of Motor Neuron 1 Protein
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Survival of Motor Neuron 2 Protein
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Green Fluorescent Proteins
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Luciferases
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Histone Acetyltransferases