title: Epidermal Growth Factor-dependent Activation of the Extracellular Signal-regulated Kinase Pathway by DJ-1 Protein through Its Direct Binding to c-Raf Protein.
abstract: DJ-1 is an oncogene and also a causative gene for familial Parkinson disease. DJ-1 has various functions, and the oxidative status of cysteine at position 106 (Cys-106) is crucial for determination of the activation level of DJ-1. Although DJ-1 requires activated Ras for its oncogenic activity and although it activates the extracellular signal-regulated kinase (ERK) pathway, a cell growth pathway downstream of Ras, the precise mechanism underlying activation of the ERK pathway by DJ-1 is still not known. In this study, we found that DJ-1 directly bound to the kinase domain of c-Raf but not to Ras and that Cys-106 mutant DJ-1 bound to c-Raf more weakly than did wild-type DJ-1. Co-localization of DJ-1 with c-Raf in the cytoplasm was enhanced in epidermal growth factor (EGF)-treated cells. Knockdown of DJ-1 expression attenuated the phosphorylation level of c-Raf in EGF-treated cells, resulting in reduced activation of MEK and ERK1/2. Although EGF-treated DJ-1 knock-out cells also showed attenuated c-Raf activation, reintroduction of wild-type DJ-1, but not C106S DJ-1, into DJ-1 knock-out cells restored c-Raf activation in a DJ-1 binding activity in a c-Raf-dependent manner. DJ-1 was not responsible for activation of c-Raf in phorbol myristate acetate-treated cells. Furthermore, DJ-1 stimulated self-phosphorylation activity of c-Raf in vitro, but DJ-1 was not a target for Raf kinase. Oxidation of Cys-106 in DJ-1 was not affected by EGF treatment. These findings showed that DJ-1 is a positive regulator of the EGF/Ras/ERK pathway through targeting c-Raf.
authors.affiliation: Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-ku, Sapporo 060-0812.
authors.forename: Kazuko
authors.initials: K
authors.lastname: Takahashi-Niki
authors.affiliation: Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-ku, Sapporo 060-0812.
authors.forename: Izumi
authors.initials: I
authors.lastname: Kato-Ose
authors.affiliation: Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-ku, Sapporo 060-0812.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Murata
authors.affiliation: Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-ku, Sapporo 060-0812.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Maita
authors.affiliation: Faculty of Agriculture, Hokkaido University, Kita-ku, Sapporo 060-8589, Japan.
authors.forename: Sanae M M
authors.initials: SMM
authors.lastname: Iguchi-Ariga
authors.affiliation: Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-ku, Sapporo 060-0812. Electronic address: hiro@pharm.hokudai.ac.jp.
authors.forename: Hiroyoshi
authors.initials: H
authors.lastname: Ariga
mesh_terms: D000818:Animals; D002460:Cell Line; D004815:Epidermal Growth Factor / Q000032:analysis / Q000378:metabolism*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis / Q000378:metabolism*; D020935:MAP Kinase Signaling System; D051379:Mice; D015513:Oncogene Proteins / Q000032:analysis / Q000378:metabolism*; D054464:Peroxiredoxins / Q000032:analysis / Q000378:metabolism; D000071617:Protein Deglycase DJ-1; D017434:Protein Structure, Tertiary; D019908:Proto-Oncogene Proteins c-raf / Q000032:analysis / Q000378:metabolism*
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D015513:Oncogene Proteins; D004815:Epidermal Growth Factor; D054464:Peroxiredoxins; D019908:Proto-Oncogene Proteins c-raf; D048049:Extracellular Signal-Regulated MAP Kinases; C105131:PARK7 protein, human; C503466:PARK7 protein, mouse; D000071617:Protein Deglycase DJ-1
keywords: Parkinson disease (autosomal recessive, early onset) 7 (PARK7); Raf kinase; extracellular-signal-regulated kinase (ERK); protein kinase; signal transduction