title: Oxidative Dimerization of PHD2 is Responsible for its Inactivation and Contributes to Metabolic Reprogramming via HIF-1α Activation.
abstract: Prolyl hydroxylase domain protein 2 (PHD2) belongs to an evolutionarily conserved superfamily of 2-oxoglutarate and Fe(II)-dependent dioxygenases that mediates homeostatic responses to oxygen deprivation by mediating hypoxia-inducible factor-1α (HIF-1α) hydroxylation and degradation. Although oxidative stress contributes to the inactivation of PHD2, the precise molecular mechanism of PHD2 inactivation independent of the levels of co-factors is not understood. Here, we identified disulfide bond-mediated PHD2 homo-dimer formation in response to oxidative stress caused by oxidizing agents and oncogenic H-ras(V12) signalling. Cysteine residues in the double-stranded β-helix fold that constitutes the catalytic site of PHD isoforms appeared responsible for the oxidative dimerization. Furthermore, we demonstrated that disulfide bond-mediated PHD2 dimerization is associated with the stabilization and activation of HIF-1α under oxidative stress. Oncogenic H-ras(V12) signalling facilitates the accumulation of HIF-1α in the nucleus and promotes aerobic glycolysis and lactate production. Moreover, oncogenic H-ras(V12) does not trigger aerobic glycolysis in antioxidant-treated or PHD2 knocked-down cells, suggesting the participation of the ROS-mediated PHD2 inactivation in the oncogenic H-ras(V12)-mediated metabolic reprogramming. We provide here a better understanding of the mechanism by which disulfide bond-mediated PHD2 dimerization and inactivation result in the activation of HIF-1α and aerobic glycolysis in response to oxidative stress.
authors.affiliation: Department of Biomedical Chemistry, College of Biomedical &Health Science, Konkuk University, Chungju 380-701, Chungbuk, Republic of Korea.
authors.forename: Gibok
authors.initials: G
authors.lastname: Lee
authors.affiliation: Department of Biotechnology, College of Biomedical &Health Science, Konkuk University, Chungju 380-701, Chungbuk, Republic of Korea.
authors.forename: Hyung-Sik
authors.initials: HS
authors.lastname: Won
authors.affiliation: Department of Food Bioscience, College of Biomedical &Health Science, Konkuk University, Chungju 380-701, Chungbuk, Republic of Korea.
authors.forename: Yoon-Mi
authors.initials: YM
authors.lastname: Lee
authors.affiliation: Department of Biotechnology, College of Biomedical &Health Science, Konkuk University, Chungju 380-701, Chungbuk, Republic of Korea.
authors.forename: Jae-Wan
authors.initials: JW
authors.lastname: Choi
authors.affiliation: Department of Biomedical Chemistry, College of Biomedical &Health Science, Konkuk University, Chungju 380-701, Chungbuk, Republic of Korea.
authors.forename: Taek-In
authors.initials: TI
authors.lastname: Oh
authors.affiliation: Department of Biotechnology, College of Biomedical &Health Science, Konkuk University, Chungju 380-701, Chungbuk, Republic of Korea.
authors.forename: Jeong-Hwa
authors.initials: JH
authors.lastname: Jang
authors.affiliation: Department of Biotechnology, College of Biomedical &Health Science, Konkuk University, Chungju 380-701, Chungbuk, Republic of Korea.
authors.forename: Dong-Kug
authors.initials: DK
authors.lastname: Choi
authors.affiliation: Department of Biomedical Chemistry, College of Biomedical &Health Science, Konkuk University, Chungju 380-701, Chungbuk, Republic of Korea.
authors.forename: Beong-Ou
authors.initials: BO
authors.lastname: Lim
authors.affiliation: Department of Biomedical Chemistry, College of Biomedical &Health Science, Konkuk University, Chungju 380-701, Chungbuk, Republic of Korea.
authors.forename: Young Jun
authors.initials: YJ
authors.lastname: Kim
authors.affiliation: Department of Pharmacology, Seoul National University, College of Medicine, Seoul 110-799, Republic of Korea.
authors.forename: Jong-Wan
authors.initials: JW
authors.lastname: Park
authors.affiliation: Department of Cancer Biology, Dana-Farber Cancer Institute; Department of Cell Biology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Pere
authors.initials: P
authors.lastname: Puigserver
authors.affiliation: Department of Biomedical Chemistry, College of Biomedical &Health Science, Konkuk University, Chungju 380-701, Chungbuk, Republic of Korea.
authors.forename: Ji-Hong
authors.initials: JH
authors.lastname: Lim
mesh_terms: D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D003553:Cystine / Q000378:metabolism; D006019:Glycolysis; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism*; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases / Q000378:metabolism*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D055503:Protein Multimerization; D055550:Protein Stability; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction
chemical_list: C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D017382:Reactive Oxygen Species; D003553:Cystine; C490190:EGLN1 protein, human; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)