Cited by 555Mitochondrial Function and PathologyAutophagy in Disease and TherapyATP Synthase and ATPases Research

Membrane topology and mitochondrial targeting of mitofusins, ubiquitous mammalian homologs of the transmembrane GTPase Fzo

Rojo, Manuel, Legros, Frédéric, Chateau, Danielle, Lombès, Anne·2002·Journal of Cell Science

Abstract

Two human Fzo-homologs, mitofusins Mfn1 and Mfn2, are shown by RT-PCR and western blot to be ubiquitous mitochondrial proteins. Protease digestion experiments reveal that Mfn2 is an outer membrane protein with N-terminal and C-terminal domains exposed towards the cytosol. The transmembrane and C-terminal domains of Mfn2 (Mfn2-TMCT) are targeted to mitochondria and deletion of these domains leads to the cytosolic localization of truncated Mfn2 (Mfn2-NT). Mfn2 is targeted to the endoplasmic reticulum or to mitochondria when the C-terminal domain is replaced by short stretches of neutral/hydrophobic (Mfn2-IYFFT) or polar/basic (Mfn2-RRD) amino acids. The coiled-coil domains of Mfn2, upstream and downstream of the transmembrane domain, are also important for mitochondrial targeting: Mfn2-mutants deleted of any of its coiled-coil domains are only partially targeted to mitochondria and significant protein amounts remain cytosolic. We show that these coiled-coil domains interact with each other: mistargeted Mfn2-NT or Mfn2-IYFFT localize to mitochondria if co-expressed with Mfn2-TMCT. This relocalization is abolished when the coiled-coil domain is deleted in any of the co-transfected molecules. We also found that Mfn2 can cluster active mitochondria in the perinuclear region independently of the cytoskeleton,bring mitochondrial membranes into close contact and modify mitochondrial structure, without disturbing the integrity of the inner and outer membrane.

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Crossref · 10.1242/jcs.115.8.1663

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Authors
Rojo, M., Legros, F., Chateau, D., & Lombès, A.
Affiliations
  • Inserm (FR)
  • INSERM U 523 — Institut de Myologie, Groupe Hospitalier Pitié-Salpêtrière, 47 boulevard de l'Hôpital, 75651 Paris Cedex 13, France
  • Present address: INSERM U 505, Centre de Recherches Biomédicales des Cordeliers, 75006 Paris, France
Corresponding author
Rojo, Manuel
Venue
Journal of Cell Science
Published
2002-04-15
Volume
115
Issue
8
Pages
1663-1674
ISSN
  • 0021-9533 (ISSN-L)
  • 1477-9137
OpenAlex
W2117041194
Cited by
555
References
45
Citations by year
2026: 3 · 2025: 9 · 2024: 28 · 2023: 18 · 2022: 18 · 2021: 24 · 2020: 29 · 2019: 32 · 2018: 28 · 2017: 23 · 2016: 29 · 2015: 21 · 2014: 18 · 2013: 24 · 2012: 33
FWCI
6.61
Type
journal article
Language
en
Open access
closed
Locations
  • Journal of Cell Science (publishedVersion)
  • PubMed (publishedVersion)
  • HighWire Press Open Archive (submittedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
  • Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › ATP Synthase and ATPases Research
Keywords
mitofusins, Mfn1, mitofusin 2, mitochondrial targeting, membrane topology, mitochondrial outer membrane, coiled-coil domain, transmembrane domain, C-terminal domain, protein localization, mitochondrial clustering, mitochondrial structure
MeSH terms
Actins, Animals, Cell Fractionation, Cytoskeleton, HeLa Cells, Humans, Immunohistochemistry, Membrane Proteins, Microtubules, Mitochondria, Phylogeny, Protein Structure, Tertiary, GTP Phosphohydrolases, Mitochondrial Proteins, Membrane Transport Proteins, Drosophila Proteins, Mitochondrial Membrane Transport Proteins

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