Analysis of the regulatory motifs in eukaryotic initiation factor 4E-binding protein 1

Vivian H.Y. Lee, Timothy Healy, Bruno D. Fonseca, Amanda Hayashi, Christopher Proud

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Mammalian target of rapamycin complex 1 (mTORC1) phosphorylates proteins such as eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) and the S6 kinases. These substrates contain short sequences, termed TOR signalling (TOS) motifs, which interact with the mTORC1 component raptor. Phosphorylation of 4E-BP1 requires an additional feature, termed the RAIP motif (Arg-Ala-Ile-Pro). We have analysed the interaction of 4E-BP1 with raptor and the amino acid residues required for functional RAIP and TOS motifs, as assessed by raptor binding and the phosphorylation of 4E-BP1 in human cells. Binding of 4E-BP1 to raptor strongly depends on an intact TOS motif, but the RAIP motif and additional C-terminal features of 4E-BP1 also contribute to this interaction. Mutational analysis of 4E-BP1 reveals that isoleucine is a key feature of the RAIP motif, that proline is also very important and that there is greater tolerance for substitution of the first two residues. Within the TOS motif, the first position (phenylalanine in the known motifs) is most critical, whereas a wider range of residues function in other positions (although an uncharged aliphatic residue is preferred at position three). These data provide important information on the structural requirements for efficient signalling downstream of mTORC1.

LanguageEnglish
Pages2185-2199
Number of pages15
JournalFEBS Journal
Volume275
Issue number9
DOIs
Publication statusPublished - May 2008
Externally publishedYes

Keywords

  • 4E-BP1
  • mTOR
  • mTORC1
  • RAIP motif
  • TOS motif

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Lee, Vivian H.Y. ; Healy, Timothy ; Fonseca, Bruno D. ; Hayashi, Amanda ; Proud, Christopher. / Analysis of the regulatory motifs in eukaryotic initiation factor 4E-binding protein 1. In: FEBS Journal. 2008 ; Vol. 275, No. 9. pp. 2185-2199.
@article{cd3fd08f2a364b3190692f4d7b8700a9,
title = "Analysis of the regulatory motifs in eukaryotic initiation factor 4E-binding protein 1",
abstract = "Mammalian target of rapamycin complex 1 (mTORC1) phosphorylates proteins such as eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) and the S6 kinases. These substrates contain short sequences, termed TOR signalling (TOS) motifs, which interact with the mTORC1 component raptor. Phosphorylation of 4E-BP1 requires an additional feature, termed the RAIP motif (Arg-Ala-Ile-Pro). We have analysed the interaction of 4E-BP1 with raptor and the amino acid residues required for functional RAIP and TOS motifs, as assessed by raptor binding and the phosphorylation of 4E-BP1 in human cells. Binding of 4E-BP1 to raptor strongly depends on an intact TOS motif, but the RAIP motif and additional C-terminal features of 4E-BP1 also contribute to this interaction. Mutational analysis of 4E-BP1 reveals that isoleucine is a key feature of the RAIP motif, that proline is also very important and that there is greater tolerance for substitution of the first two residues. Within the TOS motif, the first position (phenylalanine in the known motifs) is most critical, whereas a wider range of residues function in other positions (although an uncharged aliphatic residue is preferred at position three). These data provide important information on the structural requirements for efficient signalling downstream of mTORC1.",
keywords = "4E-BP1, mTOR, mTORC1, RAIP motif, TOS motif",
author = "Lee, {Vivian H.Y.} and Timothy Healy and Fonseca, {Bruno D.} and Amanda Hayashi and Christopher Proud",
year = "2008",
month = "5",
doi = "10.1111/j.1742-4658.2008.06372.x",
language = "English",
volume = "275",
pages = "2185--2199",
journal = "FEBS Journal",
issn = "1742-464X",
publisher = "Wiley-Blackwell",
number = "9",

}

Analysis of the regulatory motifs in eukaryotic initiation factor 4E-binding protein 1. / Lee, Vivian H.Y.; Healy, Timothy; Fonseca, Bruno D.; Hayashi, Amanda; Proud, Christopher.

In: FEBS Journal, Vol. 275, No. 9, 05.2008, p. 2185-2199.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Analysis of the regulatory motifs in eukaryotic initiation factor 4E-binding protein 1

AU - Lee, Vivian H.Y.

AU - Healy, Timothy

AU - Fonseca, Bruno D.

AU - Hayashi, Amanda

AU - Proud, Christopher

PY - 2008/5

Y1 - 2008/5

N2 - Mammalian target of rapamycin complex 1 (mTORC1) phosphorylates proteins such as eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) and the S6 kinases. These substrates contain short sequences, termed TOR signalling (TOS) motifs, which interact with the mTORC1 component raptor. Phosphorylation of 4E-BP1 requires an additional feature, termed the RAIP motif (Arg-Ala-Ile-Pro). We have analysed the interaction of 4E-BP1 with raptor and the amino acid residues required for functional RAIP and TOS motifs, as assessed by raptor binding and the phosphorylation of 4E-BP1 in human cells. Binding of 4E-BP1 to raptor strongly depends on an intact TOS motif, but the RAIP motif and additional C-terminal features of 4E-BP1 also contribute to this interaction. Mutational analysis of 4E-BP1 reveals that isoleucine is a key feature of the RAIP motif, that proline is also very important and that there is greater tolerance for substitution of the first two residues. Within the TOS motif, the first position (phenylalanine in the known motifs) is most critical, whereas a wider range of residues function in other positions (although an uncharged aliphatic residue is preferred at position three). These data provide important information on the structural requirements for efficient signalling downstream of mTORC1.

AB - Mammalian target of rapamycin complex 1 (mTORC1) phosphorylates proteins such as eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) and the S6 kinases. These substrates contain short sequences, termed TOR signalling (TOS) motifs, which interact with the mTORC1 component raptor. Phosphorylation of 4E-BP1 requires an additional feature, termed the RAIP motif (Arg-Ala-Ile-Pro). We have analysed the interaction of 4E-BP1 with raptor and the amino acid residues required for functional RAIP and TOS motifs, as assessed by raptor binding and the phosphorylation of 4E-BP1 in human cells. Binding of 4E-BP1 to raptor strongly depends on an intact TOS motif, but the RAIP motif and additional C-terminal features of 4E-BP1 also contribute to this interaction. Mutational analysis of 4E-BP1 reveals that isoleucine is a key feature of the RAIP motif, that proline is also very important and that there is greater tolerance for substitution of the first two residues. Within the TOS motif, the first position (phenylalanine in the known motifs) is most critical, whereas a wider range of residues function in other positions (although an uncharged aliphatic residue is preferred at position three). These data provide important information on the structural requirements for efficient signalling downstream of mTORC1.

KW - 4E-BP1

KW - mTOR

KW - mTORC1

KW - RAIP motif

KW - TOS motif

UR - http://www.scopus.com/inward/record.url?scp=42449136578&partnerID=8YFLogxK

U2 - 10.1111/j.1742-4658.2008.06372.x

DO - 10.1111/j.1742-4658.2008.06372.x

M3 - Article

VL - 275

SP - 2185

EP - 2199

JO - FEBS Journal

T2 - FEBS Journal

JF - FEBS Journal

SN - 1742-464X

IS - 9

ER -