Differential regulation of ERK1/2 and mTORC1 through T1R1/T1R3 in MIN6 cells

Eric M. Wauson, Marcy L. Guerra, Julia Dyachok, Kathleen McGlynn, Jennifer Giles, Elliott M. Ross, Melanie H. Cobb

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The MAPKs ERK1/2 respond to nutrients and other insulin secretagogues in pancreatic β cells and mediate nutrient-dependent insulin gene transcription. Nutrients also stimulate the mechanistic target of rapamycin complex 1 (mTORC1) to regulate protein synthesis. We showed previously that activation of both ERK1/2 and mTORC1 in the MIN6 pancreatic β cell-derived line by extracellular amino acids (AAs) is at least in part mediated by the heterodimeric T1R1/T1R3, a G protein-coupled receptor. We show here that AAs differentially activate these two signaling pathways in MIN6 cells. Pretreatment with pertussis toxin did not prevent the activation of either ERK1/2 or mTORC1 by AAs, indicating that Gi is not central to either pathway. Although glucagonlike peptide 1, an agonist for a Gs-coupled receptor, activated ERK1/2 well and mTORC1 to a small extent, AAs had no effect on cytosolic cAMP accumulation Ca2 entry is required for ERK1/2 activation by AAs but is dispensable for AA activation of mTORC1. Pretreatment with UBO-QIC, a selective Gq inhibitor, reduced the activation of ERK1/2 but had little effect on the activation of mTORC1 by AAs, suggesting a differential requirement for Gq. Inhibition of G12/13 by the overexpression of the regulator of G protein signaling domain of p115-guanine nucleotide exchange factor had no effect on mTORC1 activation by AAs, suggesting that these G proteins are also not involved. We conclude that AAs regulate ERK1/2 and mTORC1 through distinct signaling pathways.

Original languageEnglish (US)
Pages (from-to)1114-1122
Number of pages9
JournalMolecular Endocrinology
Volume29
Issue number8
DOIs
StatePublished - Jul 31 2015

Fingerprint

Aminoacylation
Amino Acids
GTP-Binding Proteins
Food
Insulin
Guanine Nucleotide Exchange Factors
mechanistic target of rapamycin complex 1
Pertussis Toxin
G-Protein-Coupled Receptors
Cell Line
Peptides
Genes
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology

Cite this

Differential regulation of ERK1/2 and mTORC1 through T1R1/T1R3 in MIN6 cells. / Wauson, Eric M.; Guerra, Marcy L.; Dyachok, Julia; McGlynn, Kathleen; Giles, Jennifer; Ross, Elliott M.; Cobb, Melanie H.

In: Molecular Endocrinology, Vol. 29, No. 8, 31.07.2015, p. 1114-1122.

Research output: Contribution to journalArticle

Wauson, Eric M. ; Guerra, Marcy L. ; Dyachok, Julia ; McGlynn, Kathleen ; Giles, Jennifer ; Ross, Elliott M. ; Cobb, Melanie H. / Differential regulation of ERK1/2 and mTORC1 through T1R1/T1R3 in MIN6 cells. In: Molecular Endocrinology. 2015 ; Vol. 29, No. 8. pp. 1114-1122.
@article{339b890210aa4f48b743c206da5c8f6d,
title = "Differential regulation of ERK1/2 and mTORC1 through T1R1/T1R3 in MIN6 cells",
abstract = "The MAPKs ERK1/2 respond to nutrients and other insulin secretagogues in pancreatic β cells and mediate nutrient-dependent insulin gene transcription. Nutrients also stimulate the mechanistic target of rapamycin complex 1 (mTORC1) to regulate protein synthesis. We showed previously that activation of both ERK1/2 and mTORC1 in the MIN6 pancreatic β cell-derived line by extracellular amino acids (AAs) is at least in part mediated by the heterodimeric T1R1/T1R3, a G protein-coupled receptor. We show here that AAs differentially activate these two signaling pathways in MIN6 cells. Pretreatment with pertussis toxin did not prevent the activation of either ERK1/2 or mTORC1 by AAs, indicating that Gi is not central to either pathway. Although glucagonlike peptide 1, an agonist for a Gs-coupled receptor, activated ERK1/2 well and mTORC1 to a small extent, AAs had no effect on cytosolic cAMP accumulation Ca2 entry is required for ERK1/2 activation by AAs but is dispensable for AA activation of mTORC1. Pretreatment with UBO-QIC, a selective Gq inhibitor, reduced the activation of ERK1/2 but had little effect on the activation of mTORC1 by AAs, suggesting a differential requirement for Gq. Inhibition of G12/13 by the overexpression of the regulator of G protein signaling domain of p115-guanine nucleotide exchange factor had no effect on mTORC1 activation by AAs, suggesting that these G proteins are also not involved. We conclude that AAs regulate ERK1/2 and mTORC1 through distinct signaling pathways.",
author = "Wauson, {Eric M.} and Guerra, {Marcy L.} and Julia Dyachok and Kathleen McGlynn and Jennifer Giles and Ross, {Elliott M.} and Cobb, {Melanie H.}",
year = "2015",
month = "7",
day = "31",
doi = "10.1210/ME.2014-1181",
language = "English (US)",
volume = "29",
pages = "1114--1122",
journal = "Molecular Endocrinology",
issn = "0888-8809",
publisher = "The Endocrine Society",
number = "8",

}

TY - JOUR

T1 - Differential regulation of ERK1/2 and mTORC1 through T1R1/T1R3 in MIN6 cells

AU - Wauson, Eric M.

AU - Guerra, Marcy L.

AU - Dyachok, Julia

AU - McGlynn, Kathleen

AU - Giles, Jennifer

AU - Ross, Elliott M.

AU - Cobb, Melanie H.

PY - 2015/7/31

Y1 - 2015/7/31

N2 - The MAPKs ERK1/2 respond to nutrients and other insulin secretagogues in pancreatic β cells and mediate nutrient-dependent insulin gene transcription. Nutrients also stimulate the mechanistic target of rapamycin complex 1 (mTORC1) to regulate protein synthesis. We showed previously that activation of both ERK1/2 and mTORC1 in the MIN6 pancreatic β cell-derived line by extracellular amino acids (AAs) is at least in part mediated by the heterodimeric T1R1/T1R3, a G protein-coupled receptor. We show here that AAs differentially activate these two signaling pathways in MIN6 cells. Pretreatment with pertussis toxin did not prevent the activation of either ERK1/2 or mTORC1 by AAs, indicating that Gi is not central to either pathway. Although glucagonlike peptide 1, an agonist for a Gs-coupled receptor, activated ERK1/2 well and mTORC1 to a small extent, AAs had no effect on cytosolic cAMP accumulation Ca2 entry is required for ERK1/2 activation by AAs but is dispensable for AA activation of mTORC1. Pretreatment with UBO-QIC, a selective Gq inhibitor, reduced the activation of ERK1/2 but had little effect on the activation of mTORC1 by AAs, suggesting a differential requirement for Gq. Inhibition of G12/13 by the overexpression of the regulator of G protein signaling domain of p115-guanine nucleotide exchange factor had no effect on mTORC1 activation by AAs, suggesting that these G proteins are also not involved. We conclude that AAs regulate ERK1/2 and mTORC1 through distinct signaling pathways.

AB - The MAPKs ERK1/2 respond to nutrients and other insulin secretagogues in pancreatic β cells and mediate nutrient-dependent insulin gene transcription. Nutrients also stimulate the mechanistic target of rapamycin complex 1 (mTORC1) to regulate protein synthesis. We showed previously that activation of both ERK1/2 and mTORC1 in the MIN6 pancreatic β cell-derived line by extracellular amino acids (AAs) is at least in part mediated by the heterodimeric T1R1/T1R3, a G protein-coupled receptor. We show here that AAs differentially activate these two signaling pathways in MIN6 cells. Pretreatment with pertussis toxin did not prevent the activation of either ERK1/2 or mTORC1 by AAs, indicating that Gi is not central to either pathway. Although glucagonlike peptide 1, an agonist for a Gs-coupled receptor, activated ERK1/2 well and mTORC1 to a small extent, AAs had no effect on cytosolic cAMP accumulation Ca2 entry is required for ERK1/2 activation by AAs but is dispensable for AA activation of mTORC1. Pretreatment with UBO-QIC, a selective Gq inhibitor, reduced the activation of ERK1/2 but had little effect on the activation of mTORC1 by AAs, suggesting a differential requirement for Gq. Inhibition of G12/13 by the overexpression of the regulator of G protein signaling domain of p115-guanine nucleotide exchange factor had no effect on mTORC1 activation by AAs, suggesting that these G proteins are also not involved. We conclude that AAs regulate ERK1/2 and mTORC1 through distinct signaling pathways.

UR - http://www.scopus.com/inward/record.url?scp=84938344757&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84938344757&partnerID=8YFLogxK

U2 - 10.1210/ME.2014-1181

DO - 10.1210/ME.2014-1181

M3 - Article

C2 - 26168033

AN - SCOPUS:84938344757

VL - 29

SP - 1114

EP - 1122

JO - Molecular Endocrinology

JF - Molecular Endocrinology

SN - 0888-8809

IS - 8

ER -