title: ASB4 is a hydroxylation substrate of FIH and promotes vascular differentiation via an oxygen-dependent mechanism.
abstract: The molecular mechanisms of endothelial differentiation into a functional vascular network are incompletely understood. To identify novel factors in endothelial development, we used a microarray screen with differentiating embryonic stem (ES) cells that identified the gene for ankyrin repeat and SOCS box protein 4 (ASB4) as the most highly differentially expressed gene in the vascular lineage during early differentiation. Like other SOCS box-containing proteins, ASB4 is the substrate recognition molecule of an elongin B/elongin C/cullin/Roc ubiquitin ligase complex that mediates the ubiquitination and degradation of substrate protein(s). High levels of ASB4 expression in the embryonic vasculature coincide with drastic increases in oxygen tension as placental blood flow is initiated. However, as vessels mature and oxygen levels stabilize, ASB4 expression is quickly downregulated, suggesting that ASB4 may function to modulate an endothelium-specific response to increasing oxygen tension. Consistent with the hypothesis that ASB4 function is regulated by oxygen concentration, ASB4 interacts with the factor inhibiting HIF1alpha (FIH) and is a substrate for FIH-mediated hydroxylation via an oxygen-dependent mechanism. Additionally, overexpression of ASB4 in ES cells promotes differentiation into the vascular lineage in an oxygen-dependent manner. We postulate that hydroxylation of ASB4 in normoxia promotes binding to and degradation of substrate protein(s) to modulate vascular differentiation.
authors.affiliation: Department of Pharmacology, University of North Carolina, Chapel Hill, North Carolina, USA.
authors.forename: James E
authors.initials: JE
authors.lastname: Ferguson
authors.forename: Yaxu
authors.initials: Y
authors.lastname: Wu
authors.forename: Kevin
authors.initials: K
authors.lastname: Smith
authors.forename: Peter
authors.initials: P
authors.lastname: Charles
authors.forename: Kyle
authors.initials: K
authors.lastname: Powers
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Cam
authors.initials: C
authors.lastname: Patterson
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D017089:Ankyrin Repeat; D001216:Asparagine / Q000378:metabolism; D001665:Binding Sites; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D002478:Cells, Cultured; D017124:Conserved Sequence; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology; D017353:Gene Deletion; D006801:Humans; D006900:Hydroxylation; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000037:antagonists & inhibitors*; D017403:In Situ Hybridization; D007668:Kidney / Q000166:cytology; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D010100:Oxygen / Q000378:metabolism*; D017354:Point Mutation; D011233:Precipitin Tests; D011485:Protein Binding; D011487:Protein Conformation; D012333:RNA, Messenger / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity; D050826:Suppressor of Cytokine Signaling Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D020798:Two-Hybrid System Techniques
chemical_list: D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012333:RNA, Messenger; C472529:SOCS4 protein, mouse; D050826:Suppressor of Cytokine Signaling Proteins; D001216:Asparagine; D010100:Oxygen