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G protein-coupled Receptor 30 (GPR30) PDZ-dependently and Constitutively Increases ERK1/2 Signaling Through Calcineurin and Kinase Suppressor of Ras 2 (KSR2)

Olde, Bjorn LU ; de Valdivia, Ernesto Gonzalez LU ; Broselid, Stefan LU ; Kahn, Robin LU and Leeb-Lundberg, Lars M. F. LU (2016) Experimental Biology 2016 In FASEB Journal 30(1 Suppl). p.518-518
Abstract
The objective of the present study was to map the mechanism whereby G protein-coupled receptor 30 (GPR30), also called G protein-coupled estrogen receptor (GPER), stimulates extracellular signal-regulated kinase (ERK)1/2 signaling. GPR30 plays important roles in cancer and cardiometabolic regulation. We showed recently that GPR30 forms a plasma membrane complex through its C-terminal type I PSD-95/Discs-large/ZO-1 homology (PDZ) motif with a membrane-associated guanylate kinase (MAGUK) and protein kinase A (PKA)-anchoring protein 5 (AKAP5), and AKAP5-anchored PKA regulatory subunit RII suppresses receptor endocytosis and enables the receptor to constitutively inhibit cAMP production. Here, we investigated if this PDZ-dependent GPR30... (More)
The objective of the present study was to map the mechanism whereby G protein-coupled receptor 30 (GPR30), also called G protein-coupled estrogen receptor (GPER), stimulates extracellular signal-regulated kinase (ERK)1/2 signaling. GPR30 plays important roles in cancer and cardiometabolic regulation. We showed recently that GPR30 forms a plasma membrane complex through its C-terminal type I PSD-95/Discs-large/ZO-1 homology (PDZ) motif with a membrane-associated guanylate kinase (MAGUK) and protein kinase A (PKA)-anchoring protein 5 (AKAP5), and AKAP5-anchored PKA regulatory subunit RII suppresses receptor endocytosis and enables the receptor to constitutively inhibit cAMP production. Here, we investigated if this PDZ-dependent GPR30 complex also regulates ERK1/2 signaling. To do so, human and mouse GPR30 were ectopically expressed in HEK293 cells and MDCK cells, and receptors and effectors were monitored by immunoblotting, immunoprecipitation, confocal immunofluorescence microscopy, split luciferase reporter techniques, and reporter gene assays. We found that GPR30 constitutively increased extracellular signal-regulated kinase (ERK) 1/2 activity in several cell systems. The response was dependent on an intact receptor PDZ motif. Furthermore, knocking down AKAP5 or inhibiting calcineurin with FK506 inhibited the receptor response. GPR30 PDZ-dependently inhibited basal NFAT signaling, consistent with the receptor favoring AKAP5-anchoring of calcineurin. The calcineurin substrate kinase suppressor of Ras 2 (KSR2), a mitogen-activated protein kinase (MAPK) scaffold, enhanced the GPR30-promoted response, also dependently on the receptor PDZ motif. GPR30 also PDZ-dependently favored a membrane KSR2 complex at the expense of the monomeric form. On the other hand, disrupting AKAP5-PKA RII interaction with St-Ht31, or inhibiting protein kinase C (PKC) with GF109203X or epidermal growth factor receptor (EGFR) tyrosine kinase with AG1478 had no effect on the GPR30-stimulated response. FK506 also increased the amount of GPR30 in the plasma membrane, thus acting opposite to St-Ht31, which increased receptor endocytosis. We conclude that the PDZ-dependent GPR30 complex with AKAP5 includes calcineurin, which favors GPR30 endocytosis and enables the receptor to constitutively increase ERK1/2 signaling through KSR2. Support or Funding Information Swedish Cancer Foundation (Less)
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organization
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Contribution to journal
publication status
published
subject
in
FASEB Journal
volume
30
issue
1 Suppl
pages
518 - 518
publisher
The Federation of American Societies for Experimental Biology
conference name
Experimental Biology 2016
conference location
San Diego, CA, United States
conference dates
2016-04-02 - 2016-04-06
ISSN
0892-6638
DOI
10.1096/fasebj.30.1_supplement.lb518
language
English
LU publication?
yes
id
f3b11ff7-9142-408a-a8db-c1ecd24441a9
date added to LUP
2019-02-08 13:23:20
date last changed
2019-02-08 13:23:20
@misc{f3b11ff7-9142-408a-a8db-c1ecd24441a9,
  abstract     = {The objective of the present study was to map the mechanism whereby G protein-coupled receptor 30 (GPR30), also called G protein-coupled estrogen receptor (GPER), stimulates extracellular signal-regulated kinase (ERK)1/2 signaling. GPR30 plays important roles in cancer and cardiometabolic regulation. We showed recently that GPR30 forms a plasma membrane complex through its C-terminal type I PSD-95/Discs-large/ZO-1 homology (PDZ) motif with a membrane-associated guanylate kinase (MAGUK) and protein kinase A (PKA)-anchoring protein 5 (AKAP5), and AKAP5-anchored PKA regulatory subunit RII suppresses receptor endocytosis and enables the receptor to constitutively inhibit cAMP production. Here, we investigated if this PDZ-dependent GPR30 complex also regulates ERK1/2 signaling. To do so, human and mouse GPR30 were ectopically expressed in HEK293 cells and MDCK cells, and receptors and effectors were monitored by immunoblotting, immunoprecipitation, confocal immunofluorescence microscopy, split luciferase reporter techniques, and reporter gene assays. We found that GPR30 constitutively increased extracellular signal-regulated kinase (ERK) 1/2 activity in several cell systems. The response was dependent on an intact receptor PDZ motif. Furthermore, knocking down AKAP5 or inhibiting calcineurin with FK506 inhibited the receptor response. GPR30 PDZ-dependently inhibited basal NFAT signaling, consistent with the receptor favoring AKAP5-anchoring of calcineurin. The calcineurin substrate kinase suppressor of Ras 2 (KSR2), a mitogen-activated protein kinase (MAPK) scaffold, enhanced the GPR30-promoted response, also dependently on the receptor PDZ motif. GPR30 also PDZ-dependently favored a membrane KSR2 complex at the expense of the monomeric form. On the other hand, disrupting AKAP5-PKA RII interaction with St-Ht31, or inhibiting protein kinase C (PKC) with GF109203X or epidermal growth factor receptor (EGFR) tyrosine kinase with AG1478 had no effect on the GPR30-stimulated response. FK506 also increased the amount of GPR30 in the plasma membrane, thus acting opposite to St-Ht31, which increased receptor endocytosis. We conclude that the PDZ-dependent GPR30 complex with AKAP5 includes calcineurin, which favors GPR30 endocytosis and enables the receptor to constitutively increase ERK1/2 signaling through KSR2. Support or Funding Information Swedish Cancer Foundation},
  author       = {Olde, Bjorn and de Valdivia, Ernesto Gonzalez and Broselid, Stefan and Kahn, Robin and Leeb-Lundberg, Lars M. F.},
  issn         = {0892-6638},
  language     = {eng},
  location     = {San Diego, CA, United States},
  month        = {04},
  note         = {Conference Abstract},
  number       = {1 Suppl},
  pages        = {518--518},
  publisher    = {The Federation of American Societies for Experimental Biology},
  series       = {FASEB Journal},
  title        = {G protein-coupled Receptor 30 (GPR30) PDZ-dependently and Constitutively Increases ERK1/2 Signaling Through Calcineurin and Kinase Suppressor of Ras 2 (KSR2)},
  url          = {http://dx.doi.org/10.1096/fasebj.30.1_supplement.lb518},
  volume       = {30},
  year         = {2016},
}