Potentiation of receptor responses induced by prolonged binding of Gα13 and leukemia-associated RhoGEF

FASEB J. 2017 Aug;31(8):3663-3676. doi: 10.1096/fj.201700026R. Epub 2017 May 2.

Abstract

Diverse cellular functions are controlled by RhoA-GTPases, which are activated by trimeric G proteins via RhoGEFs, among others. In this study, we focused on the signaling from GPCRs to RhoA via13 and leukemia-associated RhoGEF (LARG). The activation of Gα13 was elucidated in living cells with high temporal and spatial resolution by means of FRET. The inactivation after agonist withdrawal occurred in the same range (t1/2 = 25.3 ± 2.2 s; mean ± sem; n = 22) as described for other Gα proteins. The interaction of Gα13 and LARG and the thereby-induced LARG translocation to the plasma membrane were at least 1 order of magnitude more stable after agonist withdrawal, exceeding Gα13 deactivation in the absence of LARG several fold. Consequently, we observed an almost 100-fold higher agonist sensitivity of the Gα13 LARG interaction compared to the Gα13 activation in the absence of LARG.-Bodmann, E.-L., Krett, A.-L., Bünemann, M. Potentiation of receptor responses induced by prolonged binding of Gα13 and leukemia-associated RhoGEF.

Keywords: ARHGEF12; FRET; G protein; Rho; thromboxane.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins
  • Cerebral Cortex / cytology
  • GTP-Binding Protein alpha Subunits, G12-G13 / genetics
  • GTP-Binding Protein alpha Subunits, G12-G13 / metabolism*
  • HEK293 Cells
  • Humans
  • Leukemia
  • Luminescent Proteins
  • Mice
  • Neurons / metabolism
  • Plasmids
  • Protein Binding
  • Rho Guanine Nucleotide Exchange Factors / genetics
  • Rho Guanine Nucleotide Exchange Factors / metabolism*

Substances

  • Arhgef12 protein, mouse
  • Bacterial Proteins
  • Luminescent Proteins
  • Rho Guanine Nucleotide Exchange Factors
  • yellow fluorescent protein, Bacteria
  • GTP-Binding Protein alpha Subunits, G12-G13