SGEF, a RhoG guanine nucleotide exchange factor that stimulates macropinocytosis

Mol Biol Cell. 2004 Jul;15(7):3309-19. doi: 10.1091/mbc.e04-02-0146. Epub 2004 May 7.

Abstract

SGEF (SH3-containing Guanine Nucleotide Exchange Factor) is a RhoGEF of unknown function. We found the SGEF protein to be expressed in many established cell lines and highly expressed in human liver tissue. SGEF stimulated the formation of large interconnected membrane ruffles across dorsal surfaces when expressed in fibroblasts. SGEF required its proline-rich amino-terminus to generate dorsal, but not lateral, membrane ruffles and a functional SH3 domain to colocalize with filamentous actin at sites of membrane protrusion. Full-length SGEF activated RhoG, but not Rac, when expressed in fibroblasts. Further, recombinant SGEF DH/PH protein exchanged nucleotide on RhoG, but not on Rac1 or Rac3, in vitro. Scanning electron microscopy of fibroblasts demonstrated that SGEF induced dorsal ruffles that were morphologically similar to those generated by constitutively active RhoG, but not constitutively active Rac1. Transient expression of SGEF stimulated fibroblast uptake of 10-kDa dextran, a marker of macropinocytosis. This required the full-length protein and a catalytically active DH domain. Finally, activated RhoG was found to be more effective than activated Rac, and comparable to SGEF, in its ability to trigger dextran uptake. Together, this work establishes SGEF as a RhoG exchange factor and provides evidence that both SGEF and RhoG regulate membrane dynamics in promotion of macropinocytosis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Actins / analysis
  • Actins / metabolism
  • Animals
  • Cell Line
  • Fibroblasts / metabolism
  • GTP Phosphohydrolases / analysis
  • GTP Phosphohydrolases / genetics
  • GTP Phosphohydrolases / metabolism*
  • Guanine Nucleotide Exchange Factors / analysis
  • Guanine Nucleotide Exchange Factors / genetics
  • Guanine Nucleotide Exchange Factors / physiology*
  • Humans
  • Pinocytosis / genetics
  • Pinocytosis / physiology*
  • Point Mutation / genetics
  • rho GTP-Binding Proteins

Substances

  • ARHGEF26 protein, human
  • Actins
  • Guanine Nucleotide Exchange Factors
  • RHOG protein, human
  • GTP Phosphohydrolases
  • rho GTP-Binding Proteins