García-Consuegra, InésRubio, Juan CarlosNogales-Gadea, GiselaBautista, José M.Jiménez Sáiz, Sergio LorenzoCabello Sánchez, Ana BelénLucía Mulas, AlejandroAndreu, Antoni L.Arenas, JoaquínMartín, Miguel Ángel2013-11-272013-11-272009García-Consuegra, I., Rubio, J. C., Nogales-Gadea, G., Bautista, J. M., Jiménez-Sáiz, S., Cabello-Sánchez, A. B., ..., & Martín, M. A. (2009). Novel mutations in patients with McArdle disease by analysis of skeletal muscle mRNA. Journal of Medical Genetics, 46(3), 198-202.14686244http://hdl.handle.net/11268/764Objective: Methods: Results: Conclusions: To identify pathogenic mutant alleles of the PYGM gene in "genetic manifesting heterozygous" patients with McArdle disease-that is, those in whom we could only find a sole mutant allele by genomic DNA analysis.We studied four unrelated patients. PCR-RFLP, gene sequencing, and muscle cDNA analysis were performed to search for mutations in the PYGM gene. The effects of the mutations were evaluated by in silico analysis, and gene expression was assessed by real-time polymerase chain reaction (PCR).Patient 1 was a compound heterozygous for the p.G205S missense mutation and for a novel "in frame" mutation, p.Q176_M177insVQ, resulting from a retention of six nucleotides from the 3'-end sequence of intron 4. Patient 2 was heterozygous for the common nonsense mutation p.R50X, and for a 1094 bp, c.1969+214_2177+369del mutation, spanning from intron 16 to intron 17 sequences. Furthermore, mRNA expression level was dramatically reduced consistent with nonsense mediated decay. Patient 3 was heterozygous for the p.R50X substitution, and patient 4 was heterozygous for the relatively common private Spanish mutation p.W798R. These two patients harboured a heterozygous exonic synonymous variant, p.K215K. Quantification of gene transcripts in patient 3 revealed a drastic decrease in the relative expression of the gene, which strongly supports the possibility of nonsense mediated decay.Our results indicate that skeletal muscle cDNA studies in "genetic manifesting heterozygous" patients with McArdle disease are prone to identify their second mutant allele.engMutation*Glycogen Phosphorylase, Muscle Form/*GeneticsGlycogen Storage Disease Type V/*GeneticsMuscle, Skeletal/*MetabolismRna, Messenger/*AnalysisComputer SimulationGene ExpressionGlycogen Phosphorylase, Muscle Form/MetabolismGlycogen Storage Disease Type V/PathologyHeterozygoteHumansMaleMiddle AgedPolymerase Chain ReactionPolymorphism, Restriction Fragment LengthRna, Messenger/GeneticsSequence Analysis, Dnajournal article10.1136/jmg.2008.059469restricted accessFisiología humanaEnfermedad nutricional