title: Clearance of Damaged Mitochondria Through PINK1 Stabilization by JNK and ERK MAPK Signaling in Chlorpyrifos-Treated Neuroblastoma Cells.
abstract: Mitochondrial quality control and clearance of damaged mitochondria through mitophagy are important cellular activities. Studies have shown that PTEN-induced putative protein kinase 1 (PINK1) and Parkin play central roles in triggering mitophagy; however, little is known regarding the mechanism by which PINK1 modulates mitophagy in response to reactive oxygen species (ROS)-induced stress. In this study, chlorpyrifos (CPF)-induced ROS caused mitochondrial damage and subsequent engulfing of mitochondria in double-membrane autophagic vesicles, indicating that clearance of damaged mitochondria is due to mitophagy. CPF treatment resulted in PINK1 stabilization on the outer mitochondrial membrane and subsequently increased Parkin recruitment from the cytosol to the abnormal mitochondria. We found that PINK1 physically interacts with Parkin in the mitochondria of CPF-treated cells. Furthermore, a knockdown of PINK1 strongly inhibited the LC3-II protein level by blocking Parkin recruitment. This indicates that CPF-induced mitophagy is due to PINK1 stabilization in mitochondria. We observed that PINK1 stabilization was selectively regulated by ROS-mediated c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase 1/2 (ERK1/2) signaling activation but not p38 signaling. In the mitochondria of CPF-exposed cells, pretreatment with specific inhibitors of JNK and ERK1/2 significantly decreased PINK1 stabilization and Parkin recruitment and blocked the LC3-II protein level. Specifically, JNK and ERK1/2 inhibition also dramatically blocked the interaction between PINK1 and Parkin. Our results demonstrated that PINK1 regulation plays a critical role in CPF-induced mitophagy. The simple interpretation of these results is that JNK and ERK1/2 signaling regulates PINK1/Parkin-dependent mitophagy in the mitochondria of CPF-treated cells. Overall, this study proposes a novel molecular regulatory mechanism of PINK1 stabilization under CPF exposure.
authors.affiliation: Hanyang Biomedical Research Institute, Hanyang University, 133-791, Seoul, Republic of Korea.|Graduate School of Biomedical Science and Engineering, Hanyang University, 133-791, Seoul, Republic of Korea.|Department of Pharmacology, College of Medicine, Hanyang University, 133-791, Seoul, Republic of Korea.
authors.forename: Jae Hyeon
authors.initials: JH
authors.lastname: Park
authors.affiliation: Hanyang Biomedical Research Institute, Hanyang University, 133-791, Seoul, Republic of Korea.|Department of Pharmacology, College of Medicine, Hanyang University, 133-791, Seoul, Republic of Korea.
authors.forename: Juyeon
authors.initials: J
authors.lastname: Ko
authors.affiliation: Hanyang Biomedical Research Institute, Hanyang University, 133-791, Seoul, Republic of Korea.|Graduate School of Biomedical Science and Engineering, Hanyang University, 133-791, Seoul, Republic of Korea.|Department of Pharmacology, College of Medicine, Hanyang University, 133-791, Seoul, Republic of Korea.
authors.forename: Yun Sun
authors.initials: YS
authors.lastname: Park
authors.affiliation: Hanyang Biomedical Research Institute, Hanyang University, 133-791, Seoul, Republic of Korea.|Graduate School of Biomedical Science and Engineering, Hanyang University, 133-791, Seoul, Republic of Korea.
authors.forename: Jungyun
authors.initials: J
authors.lastname: Park
authors.affiliation: Hanyang Biomedical Research Institute, Hanyang University, 133-791, Seoul, Republic of Korea. jwhwang@hanyang.ac.kr.|Graduate School of Biomedical Science and Engineering, Hanyang University, 133-791, Seoul, Republic of Korea. jwhwang@hanyang.ac.kr.
authors.forename: Jungwook
authors.initials: J
authors.lastname: Hwang
authors.affiliation: Hanyang Biomedical Research Institute, Hanyang University, 133-791, Seoul, Republic of Korea. hckoh@hanyang.ac.kr.|Department of Pharmacology, College of Medicine, Hanyang University, 133-791, Seoul, Republic of Korea. hckoh@hanyang.ac.kr.
authors.forename: Hyun Chul
authors.initials: HC
authors.lastname: Koh
mesh_terms: D045744:Cell Line, Tumor; D004390:Chlorpyrifos / Q000633:toxicity*; D002800:Cholinesterase Inhibitors / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology; D011494:Protein Kinases / Q000378:metabolism*; D055550:Protein Stability / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D000099202:PTEN-Induced Putative Kinase
chemical_list: D004390:Chlorpyrifos; D002800:Cholinesterase Inhibitors; D048049:Extracellular Signal-Regulated MAP Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D011494:Protein Kinases; D017382:Reactive Oxygen Species; D000099202:PTEN-Induced Putative Kinase
keywords: ERK1/2 MAPK; JNK; Mitophagy; PINK1 stabilization; Parkin