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Gα s Directly Drives PDZ‐RhoGEF Signaling to Cdc42
Author(s) -
CastilloKauil Alejandro,
GarcíaJiménez Irving,
CervantesVillagrana Rodolfo Daniel,
AdameGarcía Sendi Rafael,
BeltránNavarro Yarely Mabell,
Gutkind J. Silvio,
ReyesCruz Guadalupe,
VázquezPrado José
Publication year - 2021
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2021.35.s1.04945
Subject(s) - cdc42 , rhoa , pdz domain , microbiology and biotechnology , receptor , biology , phosphorylation , g protein coupled receptor , g protein , gtpase activating protein , gtpase , signal transduction , actin cytoskeleton , chemistry , cytoskeleton , biochemistry , cell
G proteins stimulate RhoGEFs that control cell morphology through actin‐cytoskeleton remodeling. Gα 12 and Gα 13 interact with RH‐RhoGEFs (RGS‐homology domain RhoGEFs), driving RhoA activation. However, whether additional Gα proteins directly regulate RH‐RhoGEFs was not known. To respond this question, we first evaluated the morphological effects of constitutively active DH/PH constructs from p115RhoGEF/ARHGEF1, PDZ‐RhoGEF (PRG)/ARHGEF11, and LARG/ARHGEF12. As expected, the three constructs promoted cell contraction and activated RhoA, consistent with their known specificity. Intriguingly, PRG‐DH/PH also induced filopodia‐like cell protrusions and directly activated Cdc42. Constitutively active Gα s (Gα s Q227L) increased the ability of PRG‐DH/PH or endogenous PRG to interact with and activate Cdc42. Using a chemogenetic approach, with DREADD receptors, we demonstrated that Gs‐coupled receptors, but not those coupled to Gi or Gq, drive PRG activity towards Cdc42. Given that Gα s interacted with PRG DH/PH region, we looked for potential inhibitory effects of the minimal fragment from PRG (linker) that binds Gα s Q227L. This construct, the PRG‐linker, interfered with Gα s Q227L:PRG interaction and PRG:Cdc42 affinity. Besides, it attenuated Gα s signaling by Gs‐DREADDs and G s ‐coupled prostaglandin receptors (assessed by phosphorylation of CREB and PKA substrates). Altogether, our results demonstrate that Gα s can recognize PRG as a novel effector directing its DH/PH catalytic module to gain affinity for Cdc42.