title: FIH Regulates Cellular Metabolism through Hydroxylation of the Deubiquitinase OTUB1.
abstract: The asparagine hydroxylase, factor inhibiting HIF (FIH), confers oxygen-dependence upon the hypoxia-inducible factor (HIF), a master regulator of the cellular adaptive response to hypoxia. Studies investigating whether asparagine hydroxylation is a general regulatory oxygen-dependent modification have identified multiple non-HIF targets for FIH. However, the functional consequences of this outside of the HIF pathway remain unclear. Here, we demonstrate that the deubiquitinase ovarian tumor domain containing ubiquitin aldehyde binding protein 1 (OTUB1) is a substrate for hydroxylation by FIH on N22. Mutation of N22 leads to a profound change in the interaction of OTUB1 with proteins important in cellular metabolism. Furthermore, in cultured cells, overexpression of N22A mutant OTUB1 impairs cellular metabolic processes when compared to wild type. Based on these data, we hypothesize that OTUB1 is a target for functional hydroxylation by FIH. Additionally, we propose that our results provide new insight into the regulation of cellular energy metabolism during hypoxic stress and the potential for targeting hydroxylases for therapeutic benefit.
authors.affiliation: Systems Biology Ireland, University College Dublin, Belfield, Dublin, Ireland.|School of Medicine and Medical Science, University College Dublin, Belfield, Dublin, Ireland.|The Conway Institute, University College Dublin, Belfield, Dublin, Ireland.|Institute of Physiology and Zürich Center for Integrative Human Physiology (ZIHP), University of Zürich, Zürich, Switzerland.
authors.forename: Carsten C
authors.initials: CC
authors.lastname: Scholz
authors.affiliation: Systems Biology Ireland, University College Dublin, Belfield, Dublin, Ireland.
authors.forename: Javier
authors.initials: J
authors.lastname: Rodriguez
authors.affiliation: Institute of Physiology and Zürich Center for Integrative Human Physiology (ZIHP), University of Zürich, Zürich, Switzerland.
authors.forename: Christina
authors.initials: C
authors.lastname: Pickel
authors.affiliation: Cambridge Institute for Medical Research, Department of Medicine, University of Cambridge, Cambridge Biomedical Research Centre, Cambridge, United Kingdom.
authors.forename: Stephen
authors.initials: S
authors.lastname: Burr
authors.affiliation: School of Biological Sciences, University of Adelaide, Adelaide, South Australia, Australia.
authors.forename: Jacqueline-Alba
authors.initials: JA
authors.lastname: Fabrizio
authors.affiliation: Institute of Physiology and Zürich Center for Integrative Human Physiology (ZIHP), University of Zürich, Zürich, Switzerland.
authors.forename: Karen A
authors.initials: KA
authors.lastname: Nolan
authors.affiliation: Institute of Physiology and Zürich Center for Integrative Human Physiology (ZIHP), University of Zürich, Zürich, Switzerland.
authors.forename: Patrick
authors.initials: P
authors.lastname: Spielmann
authors.affiliation: Systems Biology Ireland, University College Dublin, Belfield, Dublin, Ireland.|The Conway Institute, University College Dublin, Belfield, Dublin, Ireland.
authors.forename: Miguel A S
authors.initials: MA
authors.lastname: Cavadas
authors.affiliation: School of Medicine and Medical Science, University College Dublin, Belfield, Dublin, Ireland.|The Conway Institute, University College Dublin, Belfield, Dublin, Ireland.
authors.forename: Bianca
authors.initials: B
authors.lastname: Crifo
authors.affiliation: School of Medicine and Medical Science, University College Dublin, Belfield, Dublin, Ireland.|The Conway Institute, University College Dublin, Belfield, Dublin, Ireland.
authors.forename: Doug N
authors.initials: DN
authors.lastname: Halligan
authors.affiliation: Cambridge Institute for Medical Research, Department of Medicine, University of Cambridge, Cambridge Biomedical Research Centre, Cambridge, United Kingdom.
authors.forename: James A
authors.initials: JA
authors.lastname: Nathan
authors.affiliation: School of Biological Sciences, University of Adelaide, Adelaide, South Australia, Australia.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Peet
authors.affiliation: Institute of Physiology and Zürich Center for Integrative Human Physiology (ZIHP), University of Zürich, Zürich, Switzerland.
authors.forename: Roland H
authors.initials: RH
authors.lastname: Wenger
authors.affiliation: Systems Biology Ireland, University College Dublin, Belfield, Dublin, Ireland.
authors.forename: Alex
authors.initials: A
authors.lastname: Von Kriegsheim
authors.affiliation: School of Medicine and Medical Science, University College Dublin, Belfield, Dublin, Ireland.|The Conway Institute, University College Dublin, Belfield, Dublin, Ireland.
authors.forename: Eoin P
authors.initials: EP
authors.lastname: Cummins
authors.affiliation: Systems Biology Ireland, University College Dublin, Belfield, Dublin, Ireland.|School of Medicine and Medical Science, University College Dublin, Belfield, Dublin, Ireland.|The Conway Institute, University College Dublin, Belfield, Dublin, Ireland.
authors.forename: Cormac T
authors.initials: CT
authors.lastname: Taylor
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D000072017:Deubiquitinating Enzymes; D004734:Energy Metabolism; D057809:HEK293 Cells; D006801:Humans; D006900:Hydroxylation; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D055550:Protein Stability; D012097:Repressor Proteins / Q000378:metabolism*
chemical_list: D012097:Repressor Proteins; D006899:Mixed Function Oxygenases; C439790:HIF1AN protein, human; D055372:AMP-Activated Protein Kinases; D000072017:Deubiquitinating Enzymes; C538826:OTUB1 protein, human; D003546:Cysteine Endopeptidases