Analysis of functional domains of rat mitochondrial Fis1, the mitochondrial fission-stimulating protein

Journal Article · Fri Jul 29 04:00:00 EDT 2005 · Biochemical and Biophysical Research Communications

DOI:https://doi.org/10.1016/j.bbrc.2005.05.154· OSTI ID:20710874

Jofuku, Akihiro ^[1]; Ishihara, Naotada ^[1]; Mihara, Katsuyoshi ^[1]

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1. Department of Molecular Biology, Graduate School of Medical Science, Kyushu University, Fukuoka 812-8582 (Japan)

In yeast, mitochondrial-fission is regulated by the cytosolic dynamin-like GTPase (Dnm1p) in conjunction with a peripheral protein, Mdv1p, and a C-tail-anchored outer membrane protein, Fis1p. In mammals, a dynamin-related protein (Drp1) and Fis1 are involved in the mitochondrial-fission reaction as Dnm1 and Fis1 orthologues, respectively. The involvement of other component(s), such as the Mdv1 homologue, and the mechanisms regulating mitochondrial-fission remain unclear. Here, we identified rat Fis1 (rFis1) and analyzed its structure-function relationship. Blue-native-polyacrylamide gel electrophoresis revealed that rFis1 formed a {approx}200-kDa complex in the outer mitochondrial membrane. Its expression in HeLa cells promoted extensive mitochondrial fragmentation, and gene knock-down by RNAi induced extension of the mitochondrial networks. Taking advantage of these properties, we analyzed functional domains of rFis1. These experiments revealed that the N-terminal and C-terminal segments are both essential for oligomeric rFis1 interaction, and the middle TPR-like domains regulate proper oligomer assembly. Any mutations that disturb the proper oligomeric assembly compromise mitochondrial division-stimulating activity of rFis1.

• OSTI ID: 20710874
• Journal Information: Biochemical and Biophysical Research Communications, Journal Name: Biochemical and Biophysical Research Communications Journal Issue: 2 Vol. 333; ISSN BBRCA9; ISSN 0006-291X
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
ELECTROPHORESIS
GELS
GENES
HELA CELLS
MEMBRANE PROTEINS
MEMBRANES
MITOCHONDRIA
MORPHOLOGY
MUTATIONS
RATS
STRUCTURE FUNCTIONS
YEASTS