Prolyl hydroxylase domain (PHD) 2 affects cell migration and F-actin formation via RhoA/rho-associated kinase-dependent cofilin phosphorylation.
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.
Vogel
Sabine
S
Department of Cardiovascular Physiology, Universitätsmedizin Göttingen, Georg-August University Göttingen, D-37073 Göttingen, Germany.
Wottawa
Marieke
M
Farhat
Katja
K
Zieseniss
Anke
A
Schnelle
Moritz
M
Le-Huu
Sinja
S
von Ahlen
Melanie
M
Malz
Cordula
C
Camenisch
Gieri
G
Katschinski
Dörthe M
DM
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*
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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