TY - JOUR
T1 - Switching heterotrimeric G protein subunits with a chemical dimerizer
AU - Putyrski, Mateusz
AU - Schultz, Carsten
N1 - Funding Information:
We are grateful to Jean-Karim Hériché (EMBL Heidelberg, Heidelberg, Germany) for his support in computational analysis of calcium oscillation patterns. We thank Tamás Balla (NIH, Bethesda, MD, USA) for rapamycin-binding domains, Kees Jalink (NCI, Amsterdam) for the Epac-based cAMP FRET sensor, Theodorus Gadella (University of Amsterdam, Amsterdam) for cDNA of Gα q , Gβ 1 , and Gγ 2, Alexandra Newton (University of California San Diego, La Jolla, CA, USA) for pmCKAR, Alen Piljić and Gregor Reither (EMBL Heidelberg) for technical advice, Heike Stichnoth (EMBL Heidelberg) for cultured cells, and Tobias Meyer (Stanford University, Stanford, CA, USA.) for critical reading of the initial manuscript. This work was supported by the Helmholtz Association (SBCancer), and the ESF and the DFG (Schu 943/8-1).
PY - 2011/9/23
Y1 - 2011/9/23
N2 - The selective manipulation of single intracellular-signaling events remains one of the key tasks when studying signaling networks. Here, we demonstrate for the first time the stimulation of FKBP fusions of various subunits of heterotrimeric G proteins by the simple addition of the chemical dimerizer rapamycin. Activation of constitutively active Gα q, but not its GDP-bound form, leads to sustained oscillations of intracellular calcium and myo-inositol 1,4,5-trisphosphate (InsP 3) levels in HEK cells, independent of the activation of endogenous Gα q, in full agreement with the InsP 3-Ca 2+ cross-coupling model of calcium oscillations. Rapamycin-induced translocation of wild-type Gα s to the plasma membrane results in elevated cAMP levels. Activation of rapamycin-inducible Gα s or Gβ 1γ 2 evokes extensive modulation of ATP-induced calcium transients. The results demonstrate that inducible heterotrimeric G protein subunits will provide ways for dissecting G protein-coupled receptor signaling.
AB - The selective manipulation of single intracellular-signaling events remains one of the key tasks when studying signaling networks. Here, we demonstrate for the first time the stimulation of FKBP fusions of various subunits of heterotrimeric G proteins by the simple addition of the chemical dimerizer rapamycin. Activation of constitutively active Gα q, but not its GDP-bound form, leads to sustained oscillations of intracellular calcium and myo-inositol 1,4,5-trisphosphate (InsP 3) levels in HEK cells, independent of the activation of endogenous Gα q, in full agreement with the InsP 3-Ca 2+ cross-coupling model of calcium oscillations. Rapamycin-induced translocation of wild-type Gα s to the plasma membrane results in elevated cAMP levels. Activation of rapamycin-inducible Gα s or Gβ 1γ 2 evokes extensive modulation of ATP-induced calcium transients. The results demonstrate that inducible heterotrimeric G protein subunits will provide ways for dissecting G protein-coupled receptor signaling.
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U2 - 10.1016/j.chembiol.2011.07.013
DO - 10.1016/j.chembiol.2011.07.013
M3 - Article
C2 - 21944751
AN - SCOPUS:80053137035
SN - 2451-9448
VL - 18
SP - 1126
EP - 1133
JO - Cell Chemical Biology
JF - Cell Chemical Biology
IS - 9
ER -