Local control of a disorder-order transition in 4E-BP1 underpins regulation of translation via eIF4E

Shirley Tait, Kaushik Dutta, David Cowburn, Jim Warwicker, Andrew J. Doig, John E G McCarthy

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The molecular mechanism underpinning regulation of eukaryotic translation initiation factor eIF4E by 4E-BP1 has remained unclear. We use isothermal calorimetry, circular dichroism, NMR, and computational modeling to analyze how the structure of the eIF4E-binding domain of 4E-BP1 determines its affinity for the dorsal face of eIF4E and thus the ability of this regulator to act as a competitive inhibitor. This work identifies the key role of solvent-facing amino acids in 4E-BP1 that are not directly engaged in interactions with eIF4E. These amino acid residues influence the propensity of the natively unfolded binding motif to fold into a conformation, including a stretch of á-helix, that is required for tight binding to eIF4E. In so doing, they contribute to a free energy landscape for 4E-BP1 folding that is poised so that phosphorylation of S65 at the C-terminal end of the helical region can modulate the propensity of folding, and thus regulate the overall free energy of 4E-BP1 binding to eIF4E, over a physiologically significant range. Thus, phosphorylation acts as an intramolecular structural modulator that biases the free energy landscape for the disorder-order transition of 4E-BP1 by destabilizing the á-helix to favor the unfolded form that cannot bind eIF4E. This type of order-disorder regulatory mechanism is likely to be relevant to other intermolecular regulatory phenomena in the cell.

Original languageEnglish (US)
Pages (from-to)17627-17632
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume107
Issue number41
DOIs
StatePublished - Oct 12 2010

Fingerprint

Phosphorylation
Eukaryotic Initiation Factors
Amino Acids
Calorimetry
Circular Dichroism

Keywords

  • Conformational change
  • Intrinsically unstructured proteins
  • mRNA cap binding
  • Posttranscriptional control

ASJC Scopus subject areas

  • General

Cite this

Local control of a disorder-order transition in 4E-BP1 underpins regulation of translation via eIF4E. / Tait, Shirley; Dutta, Kaushik; Cowburn, David; Warwicker, Jim; Doig, Andrew J.; McCarthy, John E G.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 107, No. 41, 12.10.2010, p. 17627-17632.

Research output: Contribution to journalArticle

Tait, Shirley ; Dutta, Kaushik ; Cowburn, David ; Warwicker, Jim ; Doig, Andrew J. ; McCarthy, John E G. / Local control of a disorder-order transition in 4E-BP1 underpins regulation of translation via eIF4E. In: Proceedings of the National Academy of Sciences of the United States of America. 2010 ; Vol. 107, No. 41. pp. 17627-17632.
@article{a4eec818a409477eaf904558a64f7196,
title = "Local control of a disorder-order transition in 4E-BP1 underpins regulation of translation via eIF4E",
abstract = "The molecular mechanism underpinning regulation of eukaryotic translation initiation factor eIF4E by 4E-BP1 has remained unclear. We use isothermal calorimetry, circular dichroism, NMR, and computational modeling to analyze how the structure of the eIF4E-binding domain of 4E-BP1 determines its affinity for the dorsal face of eIF4E and thus the ability of this regulator to act as a competitive inhibitor. This work identifies the key role of solvent-facing amino acids in 4E-BP1 that are not directly engaged in interactions with eIF4E. These amino acid residues influence the propensity of the natively unfolded binding motif to fold into a conformation, including a stretch of {\'a}-helix, that is required for tight binding to eIF4E. In so doing, they contribute to a free energy landscape for 4E-BP1 folding that is poised so that phosphorylation of S65 at the C-terminal end of the helical region can modulate the propensity of folding, and thus regulate the overall free energy of 4E-BP1 binding to eIF4E, over a physiologically significant range. Thus, phosphorylation acts as an intramolecular structural modulator that biases the free energy landscape for the disorder-order transition of 4E-BP1 by destabilizing the {\'a}-helix to favor the unfolded form that cannot bind eIF4E. This type of order-disorder regulatory mechanism is likely to be relevant to other intermolecular regulatory phenomena in the cell.",
keywords = "Conformational change, Intrinsically unstructured proteins, mRNA cap binding, Posttranscriptional control",
author = "Shirley Tait and Kaushik Dutta and David Cowburn and Jim Warwicker and Doig, {Andrew J.} and McCarthy, {John E G}",
year = "2010",
month = "10",
day = "12",
doi = "10.1073/pnas.1008242107",
language = "English (US)",
volume = "107",
pages = "17627--17632",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "41",

}

TY - JOUR

T1 - Local control of a disorder-order transition in 4E-BP1 underpins regulation of translation via eIF4E

AU - Tait, Shirley

AU - Dutta, Kaushik

AU - Cowburn, David

AU - Warwicker, Jim

AU - Doig, Andrew J.

AU - McCarthy, John E G

PY - 2010/10/12

Y1 - 2010/10/12

N2 - The molecular mechanism underpinning regulation of eukaryotic translation initiation factor eIF4E by 4E-BP1 has remained unclear. We use isothermal calorimetry, circular dichroism, NMR, and computational modeling to analyze how the structure of the eIF4E-binding domain of 4E-BP1 determines its affinity for the dorsal face of eIF4E and thus the ability of this regulator to act as a competitive inhibitor. This work identifies the key role of solvent-facing amino acids in 4E-BP1 that are not directly engaged in interactions with eIF4E. These amino acid residues influence the propensity of the natively unfolded binding motif to fold into a conformation, including a stretch of á-helix, that is required for tight binding to eIF4E. In so doing, they contribute to a free energy landscape for 4E-BP1 folding that is poised so that phosphorylation of S65 at the C-terminal end of the helical region can modulate the propensity of folding, and thus regulate the overall free energy of 4E-BP1 binding to eIF4E, over a physiologically significant range. Thus, phosphorylation acts as an intramolecular structural modulator that biases the free energy landscape for the disorder-order transition of 4E-BP1 by destabilizing the á-helix to favor the unfolded form that cannot bind eIF4E. This type of order-disorder regulatory mechanism is likely to be relevant to other intermolecular regulatory phenomena in the cell.

AB - The molecular mechanism underpinning regulation of eukaryotic translation initiation factor eIF4E by 4E-BP1 has remained unclear. We use isothermal calorimetry, circular dichroism, NMR, and computational modeling to analyze how the structure of the eIF4E-binding domain of 4E-BP1 determines its affinity for the dorsal face of eIF4E and thus the ability of this regulator to act as a competitive inhibitor. This work identifies the key role of solvent-facing amino acids in 4E-BP1 that are not directly engaged in interactions with eIF4E. These amino acid residues influence the propensity of the natively unfolded binding motif to fold into a conformation, including a stretch of á-helix, that is required for tight binding to eIF4E. In so doing, they contribute to a free energy landscape for 4E-BP1 folding that is poised so that phosphorylation of S65 at the C-terminal end of the helical region can modulate the propensity of folding, and thus regulate the overall free energy of 4E-BP1 binding to eIF4E, over a physiologically significant range. Thus, phosphorylation acts as an intramolecular structural modulator that biases the free energy landscape for the disorder-order transition of 4E-BP1 by destabilizing the á-helix to favor the unfolded form that cannot bind eIF4E. This type of order-disorder regulatory mechanism is likely to be relevant to other intermolecular regulatory phenomena in the cell.

KW - Conformational change

KW - Intrinsically unstructured proteins

KW - mRNA cap binding

KW - Posttranscriptional control

UR - http://www.scopus.com/inward/record.url?scp=78049268257&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78049268257&partnerID=8YFLogxK

U2 - 10.1073/pnas.1008242107

DO - 10.1073/pnas.1008242107

M3 - Article

VL - 107

SP - 17627

EP - 17632

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 41

ER -