title: Prolyl hydroxylase domain (PHD) 2 affects cell migration and F-actin formation via RhoA/rho-associated kinase-dependent cofilin phosphorylation.
abstract: Cells are responding to hypoxia via prolyl-4-hydroxylase domain (PHD) enzymes, which are responsible for oxygen-dependent hydroxylation of the hypoxia-inducible factor (HIF)-1α subunit. To gain further insight into PHD function, we generated knockdown cell models for the PHD2 isoform, which is the main isoform regulating HIF-1α hydroxylation and thus stability in normoxia. Induction of a PHD2 knockdown in tetracycline-inducible HeLa PHD2 knockdown cells resulted in increased F-actin formation as detected by phalloidin staining. A similar effect could be observed in the stably transfected PHD2 knockdown cell clones 1B6 and 3B7. F-actin is at least in part responsible for shaping cell morphology as well as regulating cell migration. Cell migration was impaired significantly as a consequence of PHD2 knockdown in a scratch assay. Mechanistically, PHD2 knockdown resulted in activation of the RhoA (Ras homolog gene family member A)/Rho-associated kinase pathway with subsequent phosphorylation of cofilin. Because cofilin phosphorylation impairs its actin-severing function, this may explain the F-actin phenotype, thereby providing a functional link between PHD2-dependent signaling and cell motility.
authors.affiliation: Department of Cardiovascular Physiology, Universitätsmedizin Göttingen, Georg-August University Göttingen, D-37073 Göttingen, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Vogel
authors.forename: Marieke
authors.initials: M
authors.lastname: Wottawa
authors.forename: Katja
authors.initials: K
authors.lastname: Farhat
authors.forename: Anke
authors.initials: A
authors.lastname: Zieseniss
authors.forename: Moritz
authors.initials: M
authors.lastname: Schnelle
authors.forename: Sinja
authors.initials: S
authors.lastname: Le-Huu
authors.forename: Melanie
authors.initials: M
authors.lastname: von Ahlen
authors.forename: Cordula
authors.initials: C
authors.lastname: Malz
authors.forename: Gieri
authors.initials: G
authors.lastname: Camenisch
authors.forename: Dörthe M
authors.initials: DM
authors.lastname: Katschinski
mesh_terms: D051339:Actin Depolymerizing Factors / Q000378:metabolism*; D000199:Actins / Q000378:metabolism*; D002465:Cell Movement; D003599:Cytoskeleton / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases; D008954:Models, Biological; D010766:Phosphorylation; D011108:Polymers / Q000737:chemistry; D011393:Procollagen-Proline Dioxygenase / Q000378:metabolism*; D011485:Protein Binding; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D054460:rho-Associated Kinases / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D051339:Actin Depolymerizing Factors; D000199:Actins; D011108:Polymers; D020033:Protein Isoforms; C066737:RHOA protein, human; C490190:EGLN1 protein, human; D011393:Procollagen-Proline Dioxygenase; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein
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