Disruption of α-actinin-integrin interactions at focal adhesions renders osteoblasts susceptible to apoptosis

Jason W. Triplett, Fredrick M. Pavalko

Research output: Contribution to journalArticlepeer-review

38 Scopus citations

Abstract

Maintenance of bone structural integrity depends in part on the rate of apoptosis of bone-forming osteoblasts. Because substrate adhesion is an important regulator of apoptosis, we have investigated the role of focal adhesions in regulating bone cell apoptosis. To test this, we expressed a truncated form of α-actinin (ROD-GFP) that competitively displaces endogenous α-actinin from focal adhesions, thus disrupting focal adhesions. Immunofluorescence and morphometric analysis of vinculin and tyrosine phosphorylation revealed that ROD-GFP expression dramatically disrupted focal adhesion organization and reduced tyrosine phosphorylation at focal adhesions. In addition, Bcl-2 protein levels were reduced in ROD-GFP-expressing cells, but caspase 3 cleavage, poly-(ADP-ribose) polymerase cleavage, histone H2A.X phosphorylation, and cytotoxicity were not increased due to ROD-GFP expression alone. Increases in both ERK and Akt phosphorylation were also observed in ROD-GFP-expressing cells, although inhibition of either ERK or Akt individually or together failed to induce apoptosis. However, we did find that ROD-GFP expression sensitized, whereas α-actinin-GFP expression protected, cells from TNF-α-induced apoptosis. Further investigation revealed that activation of TNF-α-induced survival signals, specifically Akt phosphorylation and NF-κB activation, was inhibited in ROD-GFP-expressing cells. The reduced expression of antiapoptotic Bcl-2 and inhibited survival signaling rendered ROD-GFP-expressing cells more susceptible to TNF-α-induced apoptosis. Thus we conclude that α-actinin plays a role in regulating cell survival through stabilization of focal adhesions and regulation of TNF-α-induced survival signaling.

Original languageEnglish (US)
Pages (from-to)C909-C921
JournalAmerican Journal of Physiology - Cell Physiology
Volume291
Issue number5
DOIs
StatePublished - 2006

Keywords

  • Cytoskeleton
  • Nuclear factor-κB
  • Survival
  • Tumor necrosis factor-α

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

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