Tks5 recruits AFAP-110, p190RhoGAP, and cortactin for podosome formation

Exp Cell Res. 2009 Sep 10;315(15):2581-92. doi: 10.1016/j.yexcr.2009.06.012. Epub 2009 Jun 18.


Podosome formation in vascular smooth muscle cells is characterized by the recruitment of AFAP-110, p190RhoGAP, and cortactin, which have specific roles in Src activation, local down-regulation of RhoA activity, and actin polymerization, respectively. However, the molecular mechanism that underlies their specific recruitment to podosomes remains unknown. The scaffold protein Tks5 is localized to podosomes in Src-transformed fibroblasts and in smooth muscle cells, and may serve as a specific recruiting adapter for various components during podosome formation. We show here that induced mislocalization of Tks5 to the surface of mitochondria leads to a major subcellular redistribution of AFAP-110, p190RhoGAP, and cortactin, and to inhibition of podosome formation. Analysis of a series of similarly mistargeted deletion mutants of Tks5 indicates that the fifth SH3 domain is essential for this recruitment. A Tks5 mutant lacking the PX domain also inhibits podosome formation and induces the redistribution of AFAP-110, p190RhoGAP, and cortactin to the perinuclear area. By expressing a catalytically inactive point mutant and by siRNA-mediated expression knock-down we also provide evidence that p190RhoGAP is required for podosome formation. Together our findings demonstrate that Tks5 plays a central role in the recruitment of AFAP-110, p190RhoGAP, and cortactin to drive podosome formation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Vesicular Transport / genetics
  • Adaptor Proteins, Vesicular Transport / metabolism*
  • Animals
  • Cell Surface Extensions / metabolism*
  • Cell Surface Extensions / ultrastructure
  • Cortactin / genetics
  • Cortactin / metabolism*
  • Fibroblasts / cytology
  • Fibroblasts / metabolism
  • Humans
  • Microfilament Proteins / genetics
  • Microfilament Proteins / metabolism*
  • Mitochondria / metabolism
  • Mitochondria / ultrastructure
  • Muscle, Smooth, Vascular / cytology
  • Myocytes, Smooth Muscle* / cytology
  • Myocytes, Smooth Muscle* / metabolism
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism*
  • Point Mutation
  • Protein Structure, Tertiary
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Rats
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism*


  • AFAP 110
  • Adaptor Proteins, Vesicular Transport
  • Arhgap35 protein, rat
  • Cortactin
  • Cttn protein, rat
  • Microfilament Proteins
  • Phosphoproteins
  • RNA, Small Interfering
  • Recombinant Fusion Proteins
  • Repressor Proteins
  • SH3PXD2A protein, human