Both actin and polyproline interactions of profilin-1 are required for migration, invasion and capillary morphogenesis of vascular endothelial cells.
The objective of the present study was to evaluate how different ligand interactions of profilin-1 (Pfn1), an actin-binding protein that is upregulated during capillary morphogenesis of vascular endothelial cells (VEC), contribute to migration and capillary forming ability of VEC. We adopted a knockdown-knockin experimental system to stably express either fully functional form or mutants of Pfn1 that are impaired in binding to two of its major ligands, actin (H119E mutant) and proteins containing polyproline domains (H133S mutant), in a human dermal microvascular cell line (HmVEC) against near-null endogenous Pfn1 background. We found that silencing endogenous Pfn1 expression in HmVEC leads to slower random migration, reduced velocity of membrane protrusion and a significant impairment in matrigel-induced cord formation. Only re-expression of fully functional but not any of the two ligand-binding deficient mutants of Pfn1 rescues the above defects. We further show that loss of Pfn1 expression in VEC inhibits three-dimensional capillary morphogenesis, MMP2 secretion and ECM invasion. VEC invasion through ECM is also inhibited when actin and polyproline interactions of Pfn1 are disrupted. Together, these experimental data demonstrate that Pfn1 regulates VEC migration, invasion and capillary morphogenesis through its interaction with both actin and proline-rich ligands.
Ding
Zhijie
Z
Department of Bioengineering, University of Pittsburgh, PA, USA.
Gau
David
D
Deasy
Bridget
B
Wells
Alan
A
Roy
Partha
P
D000199:Actins / Q000378:metabolism; D002196:Capillaries / Q000166:cytology* / Q000502:physiology*; D002465:Cell Movement / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D005780:Gelatin / Q000032:analysis; D020868:Gene Silencing; D006801:Humans; D009024:Morphogenesis / Q000502:physiology; D010455:Peptides / Q000378:metabolism; D010590:Phalloidine / Q000032:analysis; D051304:Profilins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D014471:Umbilical Veins / Q000166:cytology / Q000254:growth & development / Q000502:physiology
D000199:Actins; D010455:Peptides; D051304:Profilins; D010590:Phalloidine; C011083:polyproline; D005780:Gelatin