Epidermal Growth Factor-dependent Activation of the Extracellular Signal-regulated Kinase Pathway by DJ-1 Protein through Its Direct Binding to c-Raf Protein

• Published in: The Journal of biological chemistry Vol. 290; no. 29; pp. 17838 - 17847
• Main Authors: Takahashi-Niki, Kazuko, Kato-Ose, Izumi, Murata, Hiroaki, Maita, Hiroshi, Iguchi-Ariga, Sanae M.M., Ariga, Hiroyoshi
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 17.07.2015; American Society for Biochemistry and Molecular Biology
• Subjects: Animals; Cell Line; Epidermal Growth Factor - analysis; Epidermal Growth Factor - metabolism; Extracellular Signal-Regulated MAP Kinases - metabolism; extracellular-signal-regulated kinase (ERK); Humans; Intracellular Signaling Peptides and Proteins - analysis; Intracellular Signaling Peptides and Proteins - metabolism; MAP Kinase Signaling System; Mice; Oncogene Proteins - analysis; Oncogene Proteins - metabolism; Parkinson disease (autosomal recessive, early onset) 7 (PARK7); Peroxiredoxins - analysis; Peroxiredoxins - metabolism; Protein Deglycase DJ-1; protein kinase; Protein Structure, Tertiary; Proto-Oncogene Proteins c-raf - analysis; Proto-Oncogene Proteins c-raf - metabolism; Raf kinase; Signal Transduction; extracellular-signal-regulated kinase (ERK); Raf kinase; signal transduction; Parkinson disease (autosomal recessive, early onset) 7 (PARK7); protein kinase
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M115.666271

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. DJ-1 is a ras-cooperating oncogene and activates the ERK pathway. DJ-1 directly binds to the kinase domain of c-Raf to stimulate its self-phosphorylation, followed by phosphorylation of MEK and ERK1/2 in EGF-treated cells. DJ-1 activates the ERK pathway by directly binding to c-Raf but not to Ras. DJ-1 is a positive regulator for the EGF/Ras/ERK pathway.