PLoS Genetics

Publication date: 2013-01
ISSN: 1553-7404, 1553-7390 PMID: 23300487
DOI: 10.1371/journal.pgen.1003186
Publisher: Public Library of Science


Pistoni, Elena
Shiue, Lily ; Cline, Melissa S ; Bortolanza, Sergia ; Neguembor, Maria Victoria ; Xynos, Alexandros ; Ares Jr, Manuel ; Gabellini, Davide


Science & Technology, Life Sciences & Biomedicine, Genetics & Heredity, ACTIN-BUNDLING PROTEIN, MENTAL-RETARDATION, REGION GENE-1, DEVELOPMENTAL CONTROL, EXPRESSION PATTERNS, DNA REARRANGEMENTS, SKELETAL-MUSCLE, CANDIDATE GENE, STEM-CELLS, RNA, Alternative Splicing, Animals, Calpain, Cells, Cultured, Disease Models, Animal, Exons, Gene Expression Regulation, Developmental, HEK293 Cells, Humans, Mice, Muscle Development, Muscle Proteins, Muscular Dystrophy, Facioscapulohumeral, Myoblasts, Proteins, RNA Splicing Factors, RNA-Binding Proteins, Developmental Biology, 0604 Genetics


Facioscapulohumeral muscular dystrophy (FSHD) is a common muscle disease whose molecular pathogenesis remains largely unknown. Over-expression of FSHD region gene 1 (FRG1) in mice, frogs, and worms perturbs muscle development and causes FSHD-like phenotypes. FRG1 has been implicated in splicing, and we asked how splicing might be involved in FSHD by conducting a genome-wide analysis in FRG1 mice. We find that splicing perturbations parallel the responses of different muscles to FRG1 over-expression and disease progression. Interestingly, binding sites for the Rbfox family of splicing factors are over-represented in a subset of FRG1-affected splicing events. Rbfox1 knockdown, over-expression, and RNA-IP confirm that these are direct Rbfox1 targets. We find that FRG1 is associated to the Rbfox1 RNA and decreases its stability. Consistent with this, Rbfox1 expression is down-regulated in mice and cells over-expressing FRG1 as well as in FSHD patients. Among the genes affected is Calpain 3, which is mutated in limb girdle muscular dystrophy, a disease phenotypically similar to FSHD. In FRG1 mice and FSHD patients, the Calpain 3 isoform lacking exon 6 (Capn3 E6-) is increased. Finally, Rbfox1 knockdown and over-expression of Capn3 E6- inhibit muscle differentiation. Collectively, our results suggest that a component of FSHD pathogenesis may arise by over-expression of FRG1, reducing Rbfox1 levels and leading to aberrant expression of an altered Calpain 3 protein through dysregulated splicing.