Dietary methionine restriction regulates liver protein synthesis and gene expression independently of eukaryotic initiation factor 2 phosphorylation in mice

Ashley P. Pettit, William O. Jonsson, Albert R. Bargoud, Emily T. Mirek, Frederick F. Peelor, Yongping Wang, Thomas W. Gettys, Scot R. Kimball, Benjamin F. Miller, Karyn L. Hamilton, Ronald C. Wek, Tracy G. Anthony

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Background: The phosphorylation of eukaryotic initiation factor 2 (p-eIF2) during dietary amino acid insufficiency reduces protein synthesis and alters gene expression via the integrated stress response (ISR). Objective: We explored whether a Met-restricted (MR) diet activates the ISR to reduce body fat and regulate protein balance. Methods: Male and female mice aged 3-6 mo with either whole-body deletion of general control nonderepressible 2 (Gcn2) or liver-specific deletion of protein kinase R-like endoplasmic reticulum kinase (Perk) alongside wild-type or floxed control mice were fed an obesogenic diet sufficient in Met (0.86%) or an MR (0.12% Met) diet for ≤5 wk. Ala enrichment with deuterium was measured to calculate protein synthesis rates. The guanine nucleotide exchange factor activity of eIF2B was measured alongside p-eIF2 and hepatic mRNA expression levels at 2 d and 5 wk. Metabolic phenotyping was conducted at 4 wk, and body composition was measured throughout. Results were evaluated with the use of ANOVA (P < 0.05). Results: Feeding an MR diet for 2 d did not increase hepatic p-eIF2 or reduce eIF2B activity in wild-type or Gcn22/2-/- mice, yet many genes transcriptionally regulated by the ISR were altered in both strains in the same direction and amplitude. Feeding anMRdiet for 5wk increased p-eIF2 and reduced eIF2B activity inwild-type but not Gcn22/2-/- mice, yet ISR-regulated genes altered in both strains similarly. Furthermore, the MR diet reduced mixed and cytosolic but not mitochondrial protein synthesis in both the liver and skeletal muscle regardless of Gcn2 status. Despite the similarities between strains, the MR diet did not increase energy expenditure or reduce body fat in Gcn22/2-/- mice. Finally, feeding the MR diet to mice with Perk deleted in the liver increased hepatic p-eIF2 and altered body composition similar to floxed controls. Conclusions: Hepatic activation of the ISR resulting from anMR diet does not require p-eIF2. Gcn2 status influences body fat loss but not protein balance when Met is restricted.

Original languageEnglish (US)
Pages (from-to)1031-1040
Number of pages10
JournalJournal of Nutrition
Volume147
Issue number6
DOIs
StatePublished - Jun 1 2017

Fingerprint

Eukaryotic Initiation Factor-2
Methionine
Phosphorylation
Diet
Gene Expression
Liver
Proteins
Adipose Tissue
Body Composition
Guanine Nucleotide Exchange Factors
Deuterium
Mitochondrial Proteins
Endoplasmic Reticulum
Protein Kinases
Energy Metabolism
Genes
Analysis of Variance
Skeletal Muscle
Phosphotransferases
Amino Acids

All Science Journal Classification (ASJC) codes

  • Medicine (miscellaneous)
  • Nutrition and Dietetics

Keywords

  • ATF4
  • EIF2B
  • GCN2
  • Integrated stress response
  • PERK

Cite this

Pettit, Ashley P. ; Jonsson, William O. ; Bargoud, Albert R. ; Mirek, Emily T. ; Peelor, Frederick F. ; Wang, Yongping ; Gettys, Thomas W. ; Kimball, Scot R. ; Miller, Benjamin F. ; Hamilton, Karyn L. ; Wek, Ronald C. ; Anthony, Tracy G. / Dietary methionine restriction regulates liver protein synthesis and gene expression independently of eukaryotic initiation factor 2 phosphorylation in mice. In: Journal of Nutrition. 2017 ; Vol. 147, No. 6. pp. 1031-1040.
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abstract = "Background: The phosphorylation of eukaryotic initiation factor 2 (p-eIF2) during dietary amino acid insufficiency reduces protein synthesis and alters gene expression via the integrated stress response (ISR). Objective: We explored whether a Met-restricted (MR) diet activates the ISR to reduce body fat and regulate protein balance. Methods: Male and female mice aged 3-6 mo with either whole-body deletion of general control nonderepressible 2 (Gcn2) or liver-specific deletion of protein kinase R-like endoplasmic reticulum kinase (Perk) alongside wild-type or floxed control mice were fed an obesogenic diet sufficient in Met (0.86{\%}) or an MR (0.12{\%} Met) diet for ≤5 wk. Ala enrichment with deuterium was measured to calculate protein synthesis rates. The guanine nucleotide exchange factor activity of eIF2B was measured alongside p-eIF2 and hepatic mRNA expression levels at 2 d and 5 wk. Metabolic phenotyping was conducted at 4 wk, and body composition was measured throughout. Results were evaluated with the use of ANOVA (P < 0.05). Results: Feeding an MR diet for 2 d did not increase hepatic p-eIF2 or reduce eIF2B activity in wild-type or Gcn22/2-/- mice, yet many genes transcriptionally regulated by the ISR were altered in both strains in the same direction and amplitude. Feeding anMRdiet for 5wk increased p-eIF2 and reduced eIF2B activity inwild-type but not Gcn22/2-/- mice, yet ISR-regulated genes altered in both strains similarly. Furthermore, the MR diet reduced mixed and cytosolic but not mitochondrial protein synthesis in both the liver and skeletal muscle regardless of Gcn2 status. Despite the similarities between strains, the MR diet did not increase energy expenditure or reduce body fat in Gcn22/2-/- mice. Finally, feeding the MR diet to mice with Perk deleted in the liver increased hepatic p-eIF2 and altered body composition similar to floxed controls. Conclusions: Hepatic activation of the ISR resulting from anMR diet does not require p-eIF2. Gcn2 status influences body fat loss but not protein balance when Met is restricted.",
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author = "Pettit, {Ashley P.} and Jonsson, {William O.} and Bargoud, {Albert R.} and Mirek, {Emily T.} and Peelor, {Frederick F.} and Yongping Wang and Gettys, {Thomas W.} and Kimball, {Scot R.} and Miller, {Benjamin F.} and Hamilton, {Karyn L.} and Wek, {Ronald C.} and Anthony, {Tracy G.}",
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Pettit, AP, Jonsson, WO, Bargoud, AR, Mirek, ET, Peelor, FF, Wang, Y, Gettys, TW, Kimball, SR, Miller, BF, Hamilton, KL, Wek, RC & Anthony, TG 2017, 'Dietary methionine restriction regulates liver protein synthesis and gene expression independently of eukaryotic initiation factor 2 phosphorylation in mice', Journal of Nutrition, vol. 147, no. 6, pp. 1031-1040. https://doi.org/10.3945/jn.116.246710

Dietary methionine restriction regulates liver protein synthesis and gene expression independently of eukaryotic initiation factor 2 phosphorylation in mice. / Pettit, Ashley P.; Jonsson, William O.; Bargoud, Albert R.; Mirek, Emily T.; Peelor, Frederick F.; Wang, Yongping; Gettys, Thomas W.; Kimball, Scot R.; Miller, Benjamin F.; Hamilton, Karyn L.; Wek, Ronald C.; Anthony, Tracy G.

In: Journal of Nutrition, Vol. 147, No. 6, 01.06.2017, p. 1031-1040.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Dietary methionine restriction regulates liver protein synthesis and gene expression independently of eukaryotic initiation factor 2 phosphorylation in mice

AU - Pettit, Ashley P.

AU - Jonsson, William O.

AU - Bargoud, Albert R.

AU - Mirek, Emily T.

AU - Peelor, Frederick F.

AU - Wang, Yongping

AU - Gettys, Thomas W.

AU - Kimball, Scot R.

AU - Miller, Benjamin F.

AU - Hamilton, Karyn L.

AU - Wek, Ronald C.

AU - Anthony, Tracy G.

PY - 2017/6/1

Y1 - 2017/6/1

N2 - Background: The phosphorylation of eukaryotic initiation factor 2 (p-eIF2) during dietary amino acid insufficiency reduces protein synthesis and alters gene expression via the integrated stress response (ISR). Objective: We explored whether a Met-restricted (MR) diet activates the ISR to reduce body fat and regulate protein balance. Methods: Male and female mice aged 3-6 mo with either whole-body deletion of general control nonderepressible 2 (Gcn2) or liver-specific deletion of protein kinase R-like endoplasmic reticulum kinase (Perk) alongside wild-type or floxed control mice were fed an obesogenic diet sufficient in Met (0.86%) or an MR (0.12% Met) diet for ≤5 wk. Ala enrichment with deuterium was measured to calculate protein synthesis rates. The guanine nucleotide exchange factor activity of eIF2B was measured alongside p-eIF2 and hepatic mRNA expression levels at 2 d and 5 wk. Metabolic phenotyping was conducted at 4 wk, and body composition was measured throughout. Results were evaluated with the use of ANOVA (P < 0.05). Results: Feeding an MR diet for 2 d did not increase hepatic p-eIF2 or reduce eIF2B activity in wild-type or Gcn22/2-/- mice, yet many genes transcriptionally regulated by the ISR were altered in both strains in the same direction and amplitude. Feeding anMRdiet for 5wk increased p-eIF2 and reduced eIF2B activity inwild-type but not Gcn22/2-/- mice, yet ISR-regulated genes altered in both strains similarly. Furthermore, the MR diet reduced mixed and cytosolic but not mitochondrial protein synthesis in both the liver and skeletal muscle regardless of Gcn2 status. Despite the similarities between strains, the MR diet did not increase energy expenditure or reduce body fat in Gcn22/2-/- mice. Finally, feeding the MR diet to mice with Perk deleted in the liver increased hepatic p-eIF2 and altered body composition similar to floxed controls. Conclusions: Hepatic activation of the ISR resulting from anMR diet does not require p-eIF2. Gcn2 status influences body fat loss but not protein balance when Met is restricted.

AB - Background: The phosphorylation of eukaryotic initiation factor 2 (p-eIF2) during dietary amino acid insufficiency reduces protein synthesis and alters gene expression via the integrated stress response (ISR). Objective: We explored whether a Met-restricted (MR) diet activates the ISR to reduce body fat and regulate protein balance. Methods: Male and female mice aged 3-6 mo with either whole-body deletion of general control nonderepressible 2 (Gcn2) or liver-specific deletion of protein kinase R-like endoplasmic reticulum kinase (Perk) alongside wild-type or floxed control mice were fed an obesogenic diet sufficient in Met (0.86%) or an MR (0.12% Met) diet for ≤5 wk. Ala enrichment with deuterium was measured to calculate protein synthesis rates. The guanine nucleotide exchange factor activity of eIF2B was measured alongside p-eIF2 and hepatic mRNA expression levels at 2 d and 5 wk. Metabolic phenotyping was conducted at 4 wk, and body composition was measured throughout. Results were evaluated with the use of ANOVA (P < 0.05). Results: Feeding an MR diet for 2 d did not increase hepatic p-eIF2 or reduce eIF2B activity in wild-type or Gcn22/2-/- mice, yet many genes transcriptionally regulated by the ISR were altered in both strains in the same direction and amplitude. Feeding anMRdiet for 5wk increased p-eIF2 and reduced eIF2B activity inwild-type but not Gcn22/2-/- mice, yet ISR-regulated genes altered in both strains similarly. Furthermore, the MR diet reduced mixed and cytosolic but not mitochondrial protein synthesis in both the liver and skeletal muscle regardless of Gcn2 status. Despite the similarities between strains, the MR diet did not increase energy expenditure or reduce body fat in Gcn22/2-/- mice. Finally, feeding the MR diet to mice with Perk deleted in the liver increased hepatic p-eIF2 and altered body composition similar to floxed controls. Conclusions: Hepatic activation of the ISR resulting from anMR diet does not require p-eIF2. Gcn2 status influences body fat loss but not protein balance when Met is restricted.

KW - ATF4

KW - EIF2B

KW - GCN2

KW - Integrated stress response

KW - PERK

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