Two mitofusin proteins, mammalian homologues of FZO, with distinct functions are both required for mitochondrial fusion.
Mitochondria are dynamic organelles that undergo frequent fission and fusion or branching. Although these morphologic changes are considered crucial for cellular functions, the underlying mechanisms remain elusive, especially in mammalian cells. We characterized two rat mitochondrial outer membrane proteins, Mfn1 and Mfn2, with distinct tissue expressions, that are homologous to Drosophila Fzo, a GTPase involved in mitochondrial fusion. Expression of the GTPase-domain mutant of Mfn2 (Mfn2(K109T)) in HeLa cells induced mitochondrial fragmentation in which Mfn2(K109T) localized at the restricted domains. Immuno-electronmicroscopy revealed that Mfn2(K109T) was concentrated at the contact domains between adjacent mitochondria, suggesting that fusion of the outer membrane was arrested at some intermediate step. Mfn1 expression induced highly connected tubular network structures depending on the functional GTPase domain. The Mfn1-induced tubular networks were suppressed by co-expression with Mfn2. In vivo depletion of either isoform by RNA interference revealed that both are required to maintain normal mitochondrial morphology. The fusion of differentially-labeled mitochondria in HeLa cells subjected to depletion of either Mfn isoform and subsequent cell fusion by hemagglutinating virus of Japan revealed that both proteins have distinct functions in mitochondrial fusion. We conclude that the two Mfn isoforms cooperate in mitochondrial fusion in mammalian cells.
Eura
Yuka
Y
Department of Molecular Biology, Graduate School of Medical Science, Kyushu University, Fukuoka 812-8582.
Ishihara
Naotada
N
Yokota
Sadaki
S
Mihara
Katsuyoshi
K
D000818:Animals; D029721:Drosophila Proteins / Q000502:physiology; D020558:GTP Phosphohydrolases / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D008561:Membrane Fusion / Q000502:physiology; D008565:Membrane Proteins / Q000302:isolation & purification / Q000502:physiology*; D008928:Mitochondria / Q000378:metabolism / Q000502:physiology*; D024101:Mitochondrial Proteins / Q000302:isolation & purification / Q000378:metabolism* / Q000502:physiology*; D009024:Morphogenesis; D009154:Mutation; D020033:Protein Isoforms; D034741:RNA, Small Interfering; D051381:Rats; D029082:Sendai virus; D014018:Tissue Distribution; D014162:Transfection
D029721:Drosophila Proteins; D008565:Membrane Proteins; C489709:Mfn1 protein, rat; D024101:Mitochondrial Proteins; D020033:Protein Isoforms; D034741:RNA, Small Interfering; C107312:Fzo protein, Drosophila; D020558:GTP Phosphohydrolases; C488750:Mfn1 protein, mouse; C489710:Mfn2 protein, rat