title: CRIF1 deficiency induces p66shc-mediated oxidative stress and endothelial activation.
abstract: Mitochondrial dysfunction has been implicated in the pathophysiology of various cardiovascular diseases. CRIF1 is a protein present in the mitochondria associated with large mitoribosomal subunits, and CRIF1 knockdown induces mitochondrial dysfunction and promotes ROS production. p66shc is a redox enzyme implicated in mitochondrial ROS generation and translation of oxidative signals and, therefore, is a key factor for oxidative stress in endothelial cells. In this study, we investigated whether mitochondrial dysfunction induced by CRIF1 knockdown induces p66shc stimulation and plays any role in mitochondrial dysfunction-induced endothelial activation. Knockdown of CRIF1 decreased the expression of mitochondrial oxidative phosphorylation (OXPHOS) complexes I, III and IV, leading to increased mitochondrial ROS (mtROS) and hyperpolarization of the mitochondrial membrane potential. Knockdown of CRIF1 also stimulated phosphorylation of p66shc and increased cytosolic ROS in endothelial cells. Furthermore, the expression of vascular cell adhesion molecule-1 and endoplasmic reticulum stress proteins were increased upon CRIF1 knockdown in endothelial cells. However, p66shc knockdown blunted the alteration in mitochondrial dynamics and ROS production in CRIF1 knockdown endothelial cells. In addition, p66shc knockdown reduced the CRIF1 knockdown-induced increases in adhesion between monocytes and endothelial cells. Taken together, these results suggest that CRIF1 knockdown partially induces endothelial activation via increased ROS production and phosphorylation of p66shc.
authors.affiliation: Department of physiology, School of Medicine, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Harsha
authors.initials: H
authors.lastname: Nagar
authors.affiliation: Department of Endocrinology, Chungnam National University Hospital, Daejeon, Republic of Korea.
authors.forename: Saet-byel
authors.initials: SB
authors.lastname: Jung
authors.affiliation: Department of physiology, School of Medicine, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Sun Kwan
authors.initials: SK
authors.lastname: Kwon
authors.affiliation: Department of physiology, School of Medicine, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Jung-Bum
authors.initials: JB
authors.lastname: Park
authors.affiliation: Department of Endocrinology, Chungnam National University Hospital, Daejeon, Republic of Korea.
authors.forename: Minho
authors.initials: M
authors.lastname: Shong
authors.affiliation: Department of neurology, Chungnam National University Hospital, Daejeon, Republic of Korea.
authors.forename: Hee-Jung
authors.initials: HJ
authors.lastname: Song
authors.affiliation: Department of physiology, School of Medicine, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Byeong Hwa
authors.initials: BH
authors.lastname: Jeon
authors.affiliation: Division of Cardiovascular Medicine, Department of Internal Medicine, University of Iowa Carver College of Medicine, Iowa City, Iowa, United States of America.
authors.forename: Kaikobad
authors.initials: K
authors.lastname: Irani
authors.affiliation: Department of physiology, School of Medicine, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Cuk-Seong
authors.initials: CS
authors.lastname: Kim
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D018797:Cell Cycle Proteins / Q000172:deficiency* / Q000235:genetics; D059865:Endoplasmic Reticulum Stress; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D055785:Gene Knockdown Techniques; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology; D006801:Humans; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D009000:Monocytes / Q000166:cytology; D009687:Nuclear Proteins / Q000172:deficiency* / Q000235:genetics; D010085:Oxidative Phosphorylation; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism; D055765:Shc Signaling Adaptor Proteins / Q000378:metabolism*; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism
chemical_list: D018797:Cell Cycle Proteins; C527307:Crif1 protein, mouse; C476586:GADD45GIP1 protein, human; D009687:Nuclear Proteins; D017382:Reactive Oxygen Species; C528024:SHC1 protein, human; D055765:Shc Signaling Adaptor Proteins; C528034:Shc1 protein, mouse; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D019010:Vascular Cell Adhesion Molecule-1