Physical and functional interaction between dorfin and valosin-containing protein that are colocalized in ubiquitylated inclusions in neurodegenerative disorders

Shinsuke Ishigaki, Nozomi Hishikawa, Jun Ichi Niwa, Shun Ichiro Iemura, Tohru Natsume, Seiji Hori, Akira Kakizuka, Keiji Tanaka, Gen Sobue

Research output: Contribution to journalArticle

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Abstract

Dorfin, a RING-IBR type ubiquitin ligase (E3), can ubiquitylate mutant superoxide dismutase 1, the causative gene of familial amyotrophic lateral sclerosis (ALS). Dorfin is located in ubiquitylated inclusions (UBIs) in various neurodegenerative disorders, such as ALS and Parkinson's disease (PD). Here we report that Valosin-containing protein (VCP) directly binds to Dorfin and that VCP ATPase activity profoundly contributes to the E3 activity of Borfin. High through-put analysis using mass spectrometry identified VCP as a candidate of Dorfin-associated protein. Glycerol gradient centrifugation analysis showed that endogenous Dorfin consisted of a 400-600-kDa complex and was co-immunoprecipitated with endogenous VCP. In vitro experiments showed that Dorfin interacted directly with VCP through its C-terminal region. These two proteins were colocalized in aggresomes in HEK293 cells and UBIs in the affected neurons of ALS and PD. VCPK524A, a dominant negative form of VCP, reduced the E3 activity of Dorfin against mutant seperoxide dismutase 1, whereas it had no effect on the autoubiquitylation of Parkin. Our results indicate that VCPs functionally regulate Dorfin through direct interaction and that their functional interplay may be related to the process of UBI formation in neurodegenerative disorders, such as ALS or PD.

Original languageEnglish
Pages (from-to)51376-51385
Number of pages10
JournalJournal of Biological Chemistry
Volume279
Issue number49
DOIs
Publication statusPublished - Dec 3 2004
Externally publishedYes

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Neurodegenerative Diseases
Amyotrophic Lateral Sclerosis
Parkinson Disease
Ubiquitin-Protein Ligases
Centrifugation
HEK293 Cells
Glycerol
Neurons
Superoxide Dismutase
Mass spectrometry
Adenosine Triphosphatases
CDC48 protein
Mass Spectrometry
Proteins
Genes
Experiments

ASJC Scopus subject areas

  • Biochemistry

Cite this

Physical and functional interaction between dorfin and valosin-containing protein that are colocalized in ubiquitylated inclusions in neurodegenerative disorders. / Ishigaki, Shinsuke; Hishikawa, Nozomi; Niwa, Jun Ichi; Iemura, Shun Ichiro; Natsume, Tohru; Hori, Seiji; Kakizuka, Akira; Tanaka, Keiji; Sobue, Gen.

In: Journal of Biological Chemistry, Vol. 279, No. 49, 03.12.2004, p. 51376-51385.

Research output: Contribution to journalArticle

Ishigaki, Shinsuke ; Hishikawa, Nozomi ; Niwa, Jun Ichi ; Iemura, Shun Ichiro ; Natsume, Tohru ; Hori, Seiji ; Kakizuka, Akira ; Tanaka, Keiji ; Sobue, Gen. / Physical and functional interaction between dorfin and valosin-containing protein that are colocalized in ubiquitylated inclusions in neurodegenerative disorders. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 49. pp. 51376-51385.
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