Gαs directly drives PDZ-RhoGEF signaling to Cdc42
© 2020 Castillo-Kauil et al..
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 GαsQ227L's ability to guide PRG's 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.
| Errataetall: |
CommentIn: J Biol Chem. 2020 Dec 11;295(50):16929-16930. - PMID 33310745 |
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| Medienart: |
E-Artikel |
| Erscheinungsjahr: |
2020 |
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| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:295 |
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| Enthalten in: |
The Journal of biological chemistry - 295(2020), 50 vom: 11. Dez., Seite 16920-16928 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Castillo-Kauil, Alejandro [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 07.04.2021 Date Revised 29.02.2024 published: Print-Electronic CommentIn: J Biol Chem. 2020 Dec 11;295(50):16929-16930. - PMID 33310745 Citation Status MEDLINE |
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| doi: |
10.1074/jbc.AC120.015204 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM315940913 |
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| 245 | 1 | 0 | |a Gαs directly drives PDZ-RhoGEF signaling to Cdc42 |
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| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2020 Castillo-Kauil et al. | ||
| 520 | |a 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 GαsQ227L's ability to guide PRG's 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 | ||
| 650 | 4 | |a Journal Article | |
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| 650 | 4 | |a ARHGEF11 | |
| 650 | 4 | |a Cdc42 | |
| 650 | 4 | |a DH/PH catalytic module | |
| 650 | 4 | |a G protein–coupled receptor (GPCR) | |
| 650 | 4 | |a GPCR | |
| 650 | 4 | |a Galpha-s | |
| 650 | 4 | |a Gαs | |
| 650 | 4 | |a PDZ-RhoGEF | |
| 650 | 4 | |a PDZ-RhoGEF (PRG) | |
| 650 | 4 | |a Rho (Rho GTPase) | |
| 650 | 4 | |a Rho GTPases | |
| 650 | 4 | |a Rho guanine nucleotide exchange factor (RhoGEF) | |
| 650 | 4 | |a cell signaling | |
| 650 | 4 | |a guanine nucleotide exchange factor (GEF) | |
| 650 | 4 | |a heterotrimeric G protein | |
| 650 | 7 | |a ARHGEF11 protein, human |2 NLM | |
| 650 | 7 | |a Rho Guanine Nucleotide Exchange Factors |2 NLM | |
| 650 | 7 | |a GTP-Binding Protein alpha Subunits, G12-G13 |2 NLM | |
| 650 | 7 | |a EC 3.6.5.1 |2 NLM | |
| 650 | 7 | |a CDC42 protein, human |2 NLM | |
| 650 | 7 | |a EC 3.6.5.2 |2 NLM | |
| 650 | 7 | |a cdc42 GTP-Binding Protein |2 NLM | |
| 650 | 7 | |a EC 3.6.5.2 |2 NLM | |
| 700 | 1 | |a García-Jiménez, Irving |e verfasserin |4 aut | |
| 700 | 1 | |a Cervantes-Villagrana, Rodolfo Daniel |e verfasserin |4 aut | |
| 700 | 1 | |a Adame-García, Sendi Rafael |e verfasserin |4 aut | |
| 700 | 1 | |a Beltrán-Navarro, Yarely Mabell |e verfasserin |4 aut | |
| 700 | 1 | |a Gutkind, J Silvio |e verfasserin |4 aut | |
| 700 | 1 | |a Reyes-Cruz, Guadalupe |e verfasserin |4 aut | |
| 700 | 1 | |a Vázquez-Prado, José |e verfasserin |4 aut | |
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