title: The Effect of a Novel c.820C>T (Arg274Trp) Mutation in the Mitofusin 2 Gene on Fibroblast Metabolism and Clinical Manifestation in a Patient.
abstract: 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.
authors.affiliation: Molecular Biology Unit, Mossakowski Medical Research Centre, PAS, Warsaw, Poland.
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Beręsewicz
authors.affiliation: Molecular Biology Unit, Mossakowski Medical Research Centre, PAS, Warsaw, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Boratyńska-Jasińska
authors.affiliation: Department of Physics, University of Warsaw, Warsaw, Poland.
authors.forename: Łukasz
authors.initials: Ł
authors.lastname: Charzewski
authors.affiliation: Molecular Biology Unit, Mossakowski Medical Research Centre, PAS, Warsaw, Poland.
authors.forename: Maria
authors.initials: M
authors.lastname: Kawalec
authors.affiliation: Neuromuscular Unit, Mossakowski Medical Research Centre, PAS, Warsaw, Poland.
authors.forename: Dagmara
authors.initials: D
authors.lastname: Kabzińska
authors.affiliation: Neuromuscular Unit, Mossakowski Medical Research Centre, PAS, Warsaw, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Kochański
authors.affiliation: Department of Physics, University of Warsaw, Warsaw, Poland.
authors.forename: Krystiana A
authors.initials: KA
authors.lastname: Krzyśko
authors.affiliation: Molecular Biology Unit, Mossakowski Medical Research Centre, PAS, Warsaw, Poland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Zabłocka
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D001120:Arginine / Q000235:genetics; D016022:Case-Control Studies; D002478:Cells, Cultured; D002607:Charcot-Marie-Tooth Disease / Q000235:genetics* / Q000473:pathology; D005347:Fibroblasts / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000235:genetics*; D006801:Humans; D008297:Male; D024101:Mitochondrial Proteins / Q000235:genetics*; D020125:Mutation, Missense; D020641:Polymorphism, Single Nucleotide; D014364:Tryptophan / Q000235:genetics; D055815:Young Adult
chemical_list: D024101:Mitochondrial Proteins; D014364:Tryptophan; D001120:Arginine; D020558:GTP Phosphohydrolases; C470018:MFN2 protein, human