title: Prolyl-hydroxylase PHD3 interacts with pyruvate dehydrogenase (PDH)-E1β and regulates the cellular PDH activity.
abstract: Cells are frequently exposed to hypoxia in physiological and pathophysiological conditions in organisms. Control of energy metabolism is one of the critical functions of the hypoxic response. Hypoxia-Inducible Factor (HIF) is a central transcription factor that regulates the hypoxic response. HIF prolyl-hydroxylase PHDs are the enzymes that hydroxylate the α subunit of HIF and negatively regulate its expression. To further understand the physiological role of PHD3, proteomics were used to identify PHD3-interacting proteins, and pyruvate dehydrogenase (PDH)-E1β was identified as such a protein. PDH catalyzes the conversion of pyruvate to acetyl-coA, thus playing a key role in cellular energy metabolism. PDH activity was significantly decreased in PHD3-depleted MCF7 breast cancer cells and PHD3(-/-) MEFs. PHD3 depletion did not affect the expression of the PDH-E1α, E1β, and E2 subunits, or the phosphorylation status of E1α, but destabilized the PDH complex (PDC), resulting in less functional PDC. Finally, PHD3(-/-) cells were resistant to cell death in prolonged hypoxia with decreased production of ROS. Taken together, the study reveals that PHD3 regulates PDH activity in cells by physically interacting with PDC.
authors.affiliation: Oxygen Biology Laboratory, Medical Research Institute, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo 113-8510, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Kikuchi
authors.affiliation: Department of Biochemistry, School of Medicine, Keio University, Shinjuku-ku, Tokyo 160-8582, Japan; Japan Science and Technology Agency (JST), Exploratory Research for Advanced Technology (ERATO), Suematsu Gas Biology Project, Shinjuku-ku, Tokyo 160-8582, Japan.
authors.forename: Yoji Andrew
authors.initials: YA
authors.lastname: Minamishima
authors.affiliation: Oxygen Biology Laboratory, Medical Research Institute, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo 113-8510, Japan. Electronic address: nakayama.mtt@mri.tmd.ac.jp.
authors.forename: Koh
authors.initials: K
authors.lastname: Nakayama
mesh_terms: D000818:Animals; D016923:Cell Death; D015687:Cell Hypoxia; D002478:Cells, Cultured; D004795:Enzyme Stability; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D061986:MCF-7 Cells; D051379:Mice; D018345:Mice, Knockout; D011393:Procollagen-Proline Dioxygenase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D021122:Protein Subunits; D030481:Pyruvate Dehydrogenase (Lipoamide) / Q000737:chemistry / Q000378:metabolism*; D011768:Pyruvate Dehydrogenase Complex / Q000737:chemistry / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics
chemical_list: D021122:Protein Subunits; D011768:Pyruvate Dehydrogenase Complex; D034741:RNA, Small Interfering; C522722:PHD3 protein, mouse; D011393:Procollagen-Proline Dioxygenase; C488894:EGLN3 protein, human; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases; D030481:Pyruvate Dehydrogenase (Lipoamide); C111305:pyruvate dehydrogenase E1 beta subunit
keywords: Hypoxia; PDH-E1β; PHD3; Prolyl-hydroxylase; Pyruvate dehydrogenase complex