Multiple regulatory mechanisms for the Dictyostelium Roco protein GbpC

J Biol Chem. 2012 Jan 20;287(4):2749-58. doi: 10.1074/jbc.M111.315739. Epub 2011 Nov 26.

Abstract

GbpC is a multidomain Roco protein in Dictyostelium, involved in transduction of intracellular cGMP that is produced by chemotactic signals. We have shown previously that cGMP binding to GbpC induces an intramolecular signaling cascade by activating subsequently the GEF, Ras, and kinase domains. In this study, we report on the cellular localization of GbpC. In resting cells, the protein is present in the cytoplasm, but GbpC rapidly translocates to the cell boundary upon stimulation with the chemoattractant cAMP. Also, during the formation of cell-cell streams and osmotic shock, the protein localizes toward the plasma membrane and actin cytoskeleton. The translocation upon cAMP stimulation occurs downstream of heterotrimeric G proteins but is independent of guanylyl cyclases and the previously identified cGMP-induced intramolecular signaling cascade in GbpC. Mutations in the GRAM domain of GbpC lead to disturbed membrane association and inactivation of GbpC function during chemotaxis in vivo. Furthermore, we show that the GRAM domain itself associates with cellular membranes and binds various phospholipids in vitro. Together, the results show that GbpC receives multiple input signals that are both required for functional activity in vivo. cAMP-stimulation induces a cGMP-dependent signaling cascade, leading to activation of kinase activity, and, independently, cAMP induces a GRAM-dependent translocation of GbpC toward the plasma membrane and cell cortex, where it may locally phosphorylate effector proteins, which are needed for proper biological activity.

MeSH terms

  • Cell Membrane / genetics
  • Cell Membrane / metabolism*
  • Cyclic AMP / genetics
  • Cyclic AMP / metabolism*
  • Cyclic GMP / genetics
  • Cyclic GMP / metabolism*
  • Cytoplasm / genetics
  • Cytoplasm / metabolism*
  • Dictyostelium / genetics
  • Dictyostelium / metabolism*
  • Protein Structure, Tertiary
  • Protein Transport / physiology
  • Protozoan Proteins / genetics
  • Protozoan Proteins / metabolism*
  • Second Messenger Systems / physiology*

Substances

  • Protozoan Proteins
  • Cyclic AMP
  • Cyclic GMP