Poldip2 controls vascular smooth muscle cell migration by regulating focal adhesion turnover and force polarization

Srinivasa Raju Datla, Daniel J. McGrail, Sasa Vukelic, Lauren P. Huff, Alicia N. Lyle, Lily Pounkova, Minyoung Lee, Bonnie Seidel-Rogol, Mazen K. Khalil, Lula L. Hilenski, Lance S. Terada, Michelle R. Dawson, Bernard Lassègue, Kathy K. Griendling

Research output: Contribution to journalArticlepeer-review

34 Scopus citations

Abstract

Polymerase-δ-interacting protein 2 (Poldip2) interacts with NADPH oxidase 4 (Nox4) and regulates migration; however, the precise underlying mechanisms are unclear. Here, we investigated the role of Poldip2 in focal adhesion turnover, as well as traction force generation and polarization. Poldip2 overexpression (AdPoldip2) in vascular smooth muscle cells (VSMCs) impairs PDGF-induced migration and induces a characteristic phenotype of long cytoplasmic extensions. AdPoldip2 also prevents the decrease in spreading and increased aspect ratio observed in response to PDGF and slightly impairs cell contraction. Moreover, AdPoldip2 blocks focal adhesion dissolution and sustains H2O2 levels in focal adhesions, whereas Poldip2 knockdown (siPoldip2) significantly decreases the number of focal adhesions. RhoA activity is unchanged when focal adhesion dissolution is stimulated in control cells but increases in AdPoldip2-treated cells. Inhibition of RhoA blocks Poldip2-mediated attenuation of focal adhesion dissolution, and overexpression of RhoA or focal adhesion kinase (FAK) reverses the loss of focal adhesions induced by siPoldip2, indicating that RhoA and FAK mediate the effect of Poldip2 on focal adhesions. Nox4 silencing prevents focal adhesion stabilization by AdPoldip2 and induces a phenotype similar to siPoldip2, suggesting a role for Nox4 in Poldip2-induced focal adhesion stability. As a consequence of impaired focal adhesion turnover, PDGF-treated AdPoldip2 cells are unable to reduce and polarize traction forces, a necessary first step in migration. These results implicate Poldip2 in VSMC migration via regulation of focal adhesion turnover and traction force generation in a Nox4/RhoA/FAK-dependent manner.

Original languageEnglish (US)
Pages (from-to)H945-H957
JournalAmerican Journal of Physiology - Heart and Circulatory Physiology
Volume307
Issue number7
DOIs
StatePublished - Oct 1 2014
Externally publishedYes

Keywords

  • Cytoskeleton
  • Focal adhesions
  • NADPH oxidase 4
  • Polymerase-δ-interacting protein 2
  • Reactive oxygen species
  • Traction forces
  • Vascular smooth muscle cells

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

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