title: Cloning and characterization of the rat HIF-1 alpha prolyl-4-hydroxylase-1 gene.
abstract: Prolyl-4-hydroxylase domain-containing enzymes (PHDs) mediate the oxygen-dependent regulation of the heterodimeric transcription factor hypoxia-inducible factor-1 (HIF-1). Under normoxic conditions, one of the subunits of HIF-1, HIF-1alpha, is hydroxylated on specific proline residues to target HIF-1alpha for degradation by the ubiquitin-proteasome pathway. Under hypoxic conditions, the hydroxylation by the PHDs is attenuated by lack of the oxygen substrate, allowing HIF-1 to accumulate, translocate to the nucleus, and mediate HIF-mediated gene transcription. In several mammalian species including humans, three PHDs have been identified. We report here the cloning of a full-length rat cDNA that is highly homologous to the human and murine PHD-1 enzymes and encodes a protein that is 416 amino acids long. Both cDNA and protein are widely expressed in rat tissues and cell types. We demonstrate that purified and crude baculovirus-expressed rat PHD-1 exhibits HIF-1alpha specific prolyl hydroxylase activity with similar substrate affinities and is comparable to human PHD-1 protein.
authors.affiliation: Systems Biology, Berlex Biosciences, Richmond, CA 94806, USA. Ronald_Cobb@berlex.com
authors.forename: Ronald R
authors.initials: RR
authors.lastname: Cobb
authors.forename: John
authors.initials: J
authors.lastname: McClary
authors.forename: Warren
authors.initials: W
authors.lastname: Manzana
authors.forename: Silke
authors.initials: S
authors.lastname: Finster
authors.forename: Brent
authors.initials: B
authors.lastname: Larsen
authors.forename: Eric
authors.initials: E
authors.lastname: Blasko
authors.forename: Jennifer
authors.initials: J
authors.lastname: Pearson
authors.forename: Sara
authors.initials: S
authors.lastname: Biancalana
authors.forename: Katalin
authors.initials: K
authors.lastname: Kauser
authors.forename: Peter
authors.initials: P
authors.lastname: Bringmann
authors.forename: David R
authors.initials: DR
authors.lastname: Light
authors.forename: Sabine
authors.initials: S
authors.lastname: Schirm
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D006801:Humans; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics; D011393:Procollagen-Proline Dioxygenase / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D018411:Spodoptera
chemical_list: D012333:RNA, Messenger; D011393:Procollagen-Proline Dioxygenase