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

PLoS One. 2017 Jan 11;12(1):e0169999. doi: 10.1371/journal.pone.0169999. eCollection 2017.

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.

Publication types

  • Case Reports

MeSH terms

  • Amino Acid Substitution / genetics
  • Arginine / genetics
  • Case-Control Studies
  • Cells, Cultured
  • Charcot-Marie-Tooth Disease / genetics*
  • Charcot-Marie-Tooth Disease / pathology
  • Fibroblasts / metabolism
  • GTP Phosphohydrolases / genetics*
  • Humans
  • Male
  • Mitochondrial Proteins / genetics*
  • Mutation, Missense
  • Polymorphism, Single Nucleotide*
  • Tryptophan / genetics
  • Young Adult

Substances

  • Mitochondrial Proteins
  • Tryptophan
  • Arginine
  • GTP Phosphohydrolases
  • MFN2 protein, human

Supplementary concepts

  • Charcot-Marie-Tooth disease, Type 2A

Grant support

This work was supported by the National Science Centre grant NN402474640. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.