Ser-64 and Ser-111 in PHAS-I are dispensable for insulin-stimulated dissociation from eIF4E

Gail Ferguson, Isabelle Mothe-Satney, John C Lawrence

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Insulin stimulates phosphorylation of multiple sites in the eIF4E-binding protein, PHAS-I, leading to dissociation of the PHAS-I.eIF4E complex and to an increase in cap-dependent translation. The Ser-64 and Ser-111 sites have been proposed to have key roles in controlling the association of PHAS-I and eIF4E. To determine whether the effects of insulin require these sites, we assessed the control of PHAS-I proteins having Ala-64 or Ala-111 mutations. The results indicate that phosphorylation of neither site is required for insulin to promote release of PHAS-I from eIF4E. Also, the mutation of Ser-111, which has been proposed to serve as a necessary priming site for the phosphorylation of other sites in PHAS-I, did not affect the phosphorylation of Thr-36/45, Ser-64, or Thr-69. Insulin promoted the release of eIF4E from PHAS-II, a PHAS isoform that lacks the Ser-111 site, but it was without effect on the amount of eIF4E bound to the third isoform, PHAS-III. The results demonstrate that contrary to widely accepted models, Ser-64 and Ser-111 are not required for the control of PHAS-I binding to eIF4E in cells, implicating phosphorylation of the Thr sites in dissociation of the PHAS-I.eIF4E complex. The findings also indicate that PHAS-II, but not PHAS-III, contributes to the control of protein synthesis by insulin.

Original languageEnglish
Pages (from-to)47459-65
Number of pages7
JournalJournal of Biological Chemistry
Volume278
Issue number48
DOIs
Publication statusPublished - 28 Nov 2003
Externally publishedYes

Fingerprint

Phosphorylation
Insulin
Protein Isoforms
Mutation
Carrier Proteins
Proteins
Association reactions

Keywords

  • Adaptor Proteins, Signal Transducing
  • Alanine/chemistry
  • Binding Sites
  • Carrier Proteins/chemistry
  • Cell Line
  • DNA, Complementary/metabolism
  • Eukaryotic Initiation Factor-4E/chemistry
  • Eukaryotic Initiation Factors/chemistry
  • Genetic Vectors
  • Humans
  • Insulin/metabolism
  • Mutation
  • Phosphoproteins/chemistry
  • Phosphorylation
  • Precipitin Tests
  • Protein Binding
  • Protein Biosynthesis
  • Protein Isoforms
  • Protein Structure, Tertiary
  • Serine/chemistry
  • Sirolimus/pharmacology
  • Threonine/chemistry
  • Transfection

Cite this

Ferguson, Gail ; Mothe-Satney, Isabelle ; Lawrence, John C. / Ser-64 and Ser-111 in PHAS-I are dispensable for insulin-stimulated dissociation from eIF4E. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 48. pp. 47459-65.
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abstract = "Insulin stimulates phosphorylation of multiple sites in the eIF4E-binding protein, PHAS-I, leading to dissociation of the PHAS-I.eIF4E complex and to an increase in cap-dependent translation. The Ser-64 and Ser-111 sites have been proposed to have key roles in controlling the association of PHAS-I and eIF4E. To determine whether the effects of insulin require these sites, we assessed the control of PHAS-I proteins having Ala-64 or Ala-111 mutations. The results indicate that phosphorylation of neither site is required for insulin to promote release of PHAS-I from eIF4E. Also, the mutation of Ser-111, which has been proposed to serve as a necessary priming site for the phosphorylation of other sites in PHAS-I, did not affect the phosphorylation of Thr-36/45, Ser-64, or Thr-69. Insulin promoted the release of eIF4E from PHAS-II, a PHAS isoform that lacks the Ser-111 site, but it was without effect on the amount of eIF4E bound to the third isoform, PHAS-III. The results demonstrate that contrary to widely accepted models, Ser-64 and Ser-111 are not required for the control of PHAS-I binding to eIF4E in cells, implicating phosphorylation of the Thr sites in dissociation of the PHAS-I.eIF4E complex. The findings also indicate that PHAS-II, but not PHAS-III, contributes to the control of protein synthesis by insulin.",
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Ser-64 and Ser-111 in PHAS-I are dispensable for insulin-stimulated dissociation from eIF4E. / Ferguson, Gail; Mothe-Satney, Isabelle; Lawrence, John C.

In: Journal of Biological Chemistry, Vol. 278, No. 48, 28.11.2003, p. 47459-65.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Ser-64 and Ser-111 in PHAS-I are dispensable for insulin-stimulated dissociation from eIF4E

AU - Ferguson, Gail

AU - Mothe-Satney, Isabelle

AU - Lawrence, John C

PY - 2003/11/28

Y1 - 2003/11/28

N2 - Insulin stimulates phosphorylation of multiple sites in the eIF4E-binding protein, PHAS-I, leading to dissociation of the PHAS-I.eIF4E complex and to an increase in cap-dependent translation. The Ser-64 and Ser-111 sites have been proposed to have key roles in controlling the association of PHAS-I and eIF4E. To determine whether the effects of insulin require these sites, we assessed the control of PHAS-I proteins having Ala-64 or Ala-111 mutations. The results indicate that phosphorylation of neither site is required for insulin to promote release of PHAS-I from eIF4E. Also, the mutation of Ser-111, which has been proposed to serve as a necessary priming site for the phosphorylation of other sites in PHAS-I, did not affect the phosphorylation of Thr-36/45, Ser-64, or Thr-69. Insulin promoted the release of eIF4E from PHAS-II, a PHAS isoform that lacks the Ser-111 site, but it was without effect on the amount of eIF4E bound to the third isoform, PHAS-III. The results demonstrate that contrary to widely accepted models, Ser-64 and Ser-111 are not required for the control of PHAS-I binding to eIF4E in cells, implicating phosphorylation of the Thr sites in dissociation of the PHAS-I.eIF4E complex. The findings also indicate that PHAS-II, but not PHAS-III, contributes to the control of protein synthesis by insulin.

AB - Insulin stimulates phosphorylation of multiple sites in the eIF4E-binding protein, PHAS-I, leading to dissociation of the PHAS-I.eIF4E complex and to an increase in cap-dependent translation. The Ser-64 and Ser-111 sites have been proposed to have key roles in controlling the association of PHAS-I and eIF4E. To determine whether the effects of insulin require these sites, we assessed the control of PHAS-I proteins having Ala-64 or Ala-111 mutations. The results indicate that phosphorylation of neither site is required for insulin to promote release of PHAS-I from eIF4E. Also, the mutation of Ser-111, which has been proposed to serve as a necessary priming site for the phosphorylation of other sites in PHAS-I, did not affect the phosphorylation of Thr-36/45, Ser-64, or Thr-69. Insulin promoted the release of eIF4E from PHAS-II, a PHAS isoform that lacks the Ser-111 site, but it was without effect on the amount of eIF4E bound to the third isoform, PHAS-III. The results demonstrate that contrary to widely accepted models, Ser-64 and Ser-111 are not required for the control of PHAS-I binding to eIF4E in cells, implicating phosphorylation of the Thr sites in dissociation of the PHAS-I.eIF4E complex. The findings also indicate that PHAS-II, but not PHAS-III, contributes to the control of protein synthesis by insulin.

KW - Adaptor Proteins, Signal Transducing

KW - Alanine/chemistry

KW - Binding Sites

KW - Carrier Proteins/chemistry

KW - Cell Line

KW - DNA, Complementary/metabolism

KW - Eukaryotic Initiation Factor-4E/chemistry

KW - Eukaryotic Initiation Factors/chemistry

KW - Genetic Vectors

KW - Humans

KW - Insulin/metabolism

KW - Mutation

KW - Phosphoproteins/chemistry

KW - Phosphorylation

KW - Precipitin Tests

KW - Protein Binding

KW - Protein Biosynthesis

KW - Protein Isoforms

KW - Protein Structure, Tertiary

KW - Serine/chemistry

KW - Sirolimus/pharmacology

KW - Threonine/chemistry

KW - Transfection

U2 - 10.1074/jbc.M307949200

DO - 10.1074/jbc.M307949200

M3 - Article

C2 - 14507920

VL - 278

SP - 47459

EP - 47465

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 48

ER -