title: Activated mitofusin 2 signals mitochondrial fusion, interferes with Bax activation, and reduces susceptibility to radical induced depolarization.
abstract: Mitochondrial fusion in higher eukaryotes requires at least two essential GTPases, Mitofusin 1 and Mitofusin 2 (Mfn2). We have created an activated mutant of Mfn2, which shows increased rates of nucleotide exchange and decreased rates of hydrolysis relative to wild type Mfn2. Mitochondrial fusion is stimulated dramatically within heterokaryons expressing this mutant, demonstrating that hydrolysis is not requisite for the fusion event, and supporting a role for Mfn2 as a signaling GTPase. Although steady-state mitochondrial fusion required the conserved intermembrane space tryptophan residue, this requirement was overcome within the context of the hydrolysis-deficient mutant. Furthermore, the punctate localization of Mfn2 is lost in the dominant active mutants, indicating that these sites are functionally controlled by changes in the nucleotide state of Mfn2. Upon staurosporine-stimulated cell death, activated Bax is recruited to the Mfn2-containing puncta; however, Bax activation and cytochrome c release are inhibited in the presence of the dominant active mutants of Mfn2. The dominant active form of Mfn2 also protected the mitochondria against free radical-induced permeability transition. In contrast to staurosporine-induced outer membrane permeability transition, pore opening induced through the introduction of free radicals was dependent upon the conserved intermembrane space residue. This is the first evidence that Mfn2 is a signaling GTPase regulating mitochondrial fusion and that the nucleotide-dependent activation of Mfn2 concomitantly protects the organelle from permeability transition. The data provide new insights into the critical relationship between mitochondrial membrane dynamics and programmed cell death.
authors.affiliation: University of Ottawa Heart Institute, Ottawa, Ontario K1Y 4W7, Canada.
authors.forename: Margaret
authors.initials: M
authors.lastname: Neuspiel
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Zunino
authors.forename: Sandhya
authors.initials: S
authors.lastname: Gangaraju
authors.forename: Peter
authors.initials: P
authors.lastname: Rippstein
authors.forename: Heidi
authors.initials: H
authors.lastname: McBride
mesh_terms: D000818:Animals; D017209:Apoptosis; D001665:Binding Sites; D019556:COS Cells; D045304:Cytochromes c / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D005456:Fluorescent Dyes / Q000494:pharmacology; D005609:Free Radicals; D020558:GTP Phosphohydrolases / Q000378:metabolism; D005799:Genes, Dominant; D006160:Guanosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000737:chemistry* / Q000378:metabolism; D009154:Mutation; D011485:Protein Binding; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism*; D019311:Staurosporine / Q000494:pharmacology; D013997:Time Factors; D014162:Transfection; D051028:bcl-2-Associated X Protein
chemical_list: C494826:BAX protein, human; D018076:DNA, Complementary; D005456:Fluorescent Dyes; D005609:Free Radicals; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D006160:Guanosine Triphosphate; D045304:Cytochromes c; D020558:GTP Phosphohydrolases; C470018:MFN2 protein, human; D019311:Staurosporine