Gαs directly drives PDZ-RhoGEF signaling to Cdc42.
In: Journal of Biological Chemistry, Jg. 295 (2020-12-11), Heft 50
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Gα proteins promote dynamic adjustments of cell shape directed by actin-cytoskeleton reorganization via their respective RhoGEF effectors. For example, Gα13 binding to the RGS-homology (RH) domains of several RH-RhoGEFs allosterically activates these proteins, causing them to expose their catalytic Dbl-homology (DH)/pleckstrin-homology (PH) regions, which triggers downstream signals. However, whether additional Gα proteins might directly regulate the RH-RhoGEFs was not known. To explore this question, we first examined the morphological effects of expressing shortened RH-RhoGEF DH/PH constructs of p115RhoGEF/ARHGEF1, PDZ-RhoGEF (PRG)/ARHGEF11, and LARG/ARHGEF12. As expected, the three constructs promoted cell contraction and activated RhoA, known to be downstream of Gα13 Intriguingly, PRG DH/PH also induced filopodia-like cell protrusions and activated Cdc42. This pathway was stimulated by constitutively active Gαs (GαsQ227L), which enabled endogenous PRG to gain affinity for Cdc42. A chemogenetic approach revealed that signaling by Gs-coupled receptors, but not by those coupled to Gi or Gq, enabled PRG to bind Cdc42. This receptor-dependent effect, as well as CREB phosphorylation, was blocked by a construct derived from the PRG:Gαs-binding region, PRG-linker. Active Gαs interacted with isolated PRG DH and PH domains and their linker. In addition, this construct interfered with GαsQ227Ls ability to guide PRGs interaction with Cdc42. Endogenous Gs-coupled prostaglandin receptors stimulated PRG binding to membrane fractions and activated signaling to PKA, and this canonical endogenous pathway was attenuated by PRG-linker. Altogether, our results demonstrate that active Gαs can recognize PRG as a novel effector directing its DH/PH catalytic module to gain affinity for Cdc42.
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Gαs directly drives PDZ-RhoGEF signaling to Cdc42.
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Autor/in / Beteiligte Person: | Castillo-Kauil, Alejandro ; García-Jiménez, Irving ; Cervantes-Villagrana, Rodolfo ; Adame-García, Sendi ; Beltrán-Navarro, Yarely ; Gutkind, J ; Reyes-Cruz, Guadalupe ; Vázquez-Prado, José |
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Zeitschrift: | Journal of Biological Chemistry, Jg. 295 (2020-12-11), Heft 50 |
Veröffentlichung: | eScholarship, University of California, 2020 |
Medientyp: | academicJournal |
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