RhoJ interacts with the GIT-PIX complex and regulates focal adhesion disassembly

Eleanor Wilson, Katarzyna Leszczynska, Natalie S. Poulter, Francesca Edelmann, Victoria A. Salisbury, Peter J. Noy, Andrea Bacon, Joshua Z. Rappoport, John K. Heath, Roy Bicknell, Victoria L. Heath*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

RhoJ is a Rho GTPase expressed in endothelial cells and tumour cells, which regulates cell motility, invasion, endothelial tube formation and focal adhesion numbers. This study aimed to further delineate the molecular function of RhoJ. Using timelapse microscopy RhoJ was found to regulate focal adhesion disassembly; small interfering RNA (siRNA)-mediated knockdown of RhoJ increased focal adhesion disassembly time, whereas expression of an active mutant (daRhoJ) decreased it. Furthermore, daRhoJ co-precipitated with the GIT-PIX complex, a regulator of focal adhesion disassembly. An interaction between daRhoJ and GIT1 was confirmed using yeast two-hybrid experiments, and this depended on the Spa homology domain of GIT1. GIT1, GIT2, b-PIX (also known as ARHGEF7) and RhoJ all colocalised in focal adhesions and depended on each other for their recruitment to focal adhesions. Functionally, the GIT-PIX complex regulated endothelial tube formation, with knockdown of both GIT1 and GIT2, or ß-PIX phenocopying RhoJ knockdown. RhoJ-knockout mice showed reduced tumour growth and diminished tumour vessel density, identifying a role for RhoJ in mediating tumour angiogenesis. These studies give new insight into the molecular function of RhoJ in regulating cell motility and tumour vessel formation.

Original languageEnglish (US)
Pages (from-to)3039-3051
Number of pages13
JournalJournal of cell science
Volume127
Issue number14
DOIs
StatePublished - 2014

Keywords

  • Angiogenesis
  • Focal adhesion
  • GIT
  • PIX
  • RhoJ

ASJC Scopus subject areas

  • Cell Biology

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