The Effect of a Novel c.820C>T (Arg274Trp) Mutation in the Mitofusin 2 Gene on Fibroblast Metabolism and Clinical Manifestation in a Patient

• Published in: PloS one 2017-01, Vol.12 (1), p.e0169999
• Main Authors: Beręsewicz, Małgorzata, Boratyńska-Jasińska, Anna, Charzewski, Łukasz, Kawalec, Maria, Kabzińska, Dagmara, Kochański, Andrzej, Krzyśko, Krystiana A, Zabłocka, Barbara
• Format: Article
• Language: English
• Subjects: Amino Acid Substitution - genetics; Arginine - genetics; Biology and Life Sciences; Case-Control Studies; Cells, Cultured; Charcot-Marie-Tooth disease; Charcot-Marie-Tooth Disease - genetics; Charcot-Marie-Tooth Disease - pathology; Deoxyribonucleic acid; DNA; Endoplasmic reticulum; Fibroblasts; Fibroblasts - metabolism; Gene expression; Genomes; GTP; GTP Phosphohydrolases - genetics; Guanosine triphosphatases; Guanosine triphosphate; Guanosinetriphosphatase; Humans; Intellectual disabilities; Male; Malfunctions; Medical research; Medicine and Health Sciences; Metabolism; Mitochondria; Mitochondrial DNA; Mitochondrial Proteins - genetics; Molecular biology; Mutation; Mutation, Missense; Neurology; Peripheral neuropathy; Physical Sciences; Polymorphism, Single Nucleotide; Protein interaction; Proteins; Tryptophan - genetics; Young Adult
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0169999

Charcot-Marie-Tooth disease type 2A (CMT2A) is an autosomal dominant axonal peripheral neuropathy caused by mutations in the mitofusin 2 gene (MFN2). Mitofusin 2 is a GTPase protein present in the outer mitochondrial membrane and responsible for regulation of mitochondrial network architecture via the fusion of mitochondria. As that fusion process is known to be strongly dependent on the GTPase activity of mitofusin 2, it is postulated that the MFN2 mutation within the GTPase domain may lead to impaired GTPase activity, and in turn to mitochondrial dysfunction. The work described here has therefore sought to verify the effects of MFN2 mutation within its GTPase domain on mitochondrial and endoplasmic reticulum morphology, as well as the mtDNA content in a cultured primary fibroblast obtained from a CMT2A patient harboring a de novo Arg274Trp mutation. In fact, all the parameters studied were affected significantly by the presence of the mutant MFN2 protein. However, using the stable model for mitofusin 2 obtained by us, we were next able to determine that the Arg274Trp mutation does not impact directly upon GTP binding. Such results were also confirmed for GTP-hydrolysis activity of MFN2 protein in patient fibroblast. We therefore suggest that the biological malfunctions observable with the disease are not consequences of impaired GTPase activity, but rather reflect an impaired contribution of the GTPase domain to other MFN2 activities involving that region, for example protein-protein interactions.