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Mitochondria undergo fusion and fission continuously to form a dynamic interconnectingnetwork.The equilibrium between fusion and fission is vital for the maintenance of mitochondrialintegrity and distribution.Although several proteins responsible for mitochondrial fission havebeen identified in mammals,the mechanism of the fission machinery remains to be clarified.Inthis study,through differential deletion experiments,we demonstrate that the N-terminal region ofMff is necessary for mitochondrial fission,whereas the intact C-terminal structure of Mff is bothnecessary and sufficient to target mitochondria.