Controlling the DNA binding specificity of bHLH proteins through intramolecular interactions

Elizebeth C Turner; Charlotte H Cureton; Chris J Weston; Oliver S Smart; Rudolf K Allemann

doi:10.1016/j.chembiol.2003.12.015

Controlling the DNA binding specificity of bHLH proteins through intramolecular interactions

Elizebeth C Turner, Charlotte H Cureton, Chris J Weston, Oliver S Smart, Rudolf K Allemann

Research output: Contribution to journal › Article › peer-review

17 Citations (Scopus)

Abstract

Reversible control of the conformation of proteins was employed to probe the relationship between flexibility and specificity of the basic helix-loop-helix protein MyoD. A fusion protein (apaMyoD) was designed where the basic DNA binding helix of MyoD was stablized by an amino-terminal extension with a sequence derived from the bee venom peptide apamin. The disulfide-stabilized helix from apamin served as a nucleus for a helix that extended for a further ten residues, thereby holding apaMyoD's DNA recognition helix in a predominantly alpha-helical conformation. The thermal stability of the DNA complexes of apaMyoD was increased by 13 degrees C relative to MyoD-bHLH. Measurements of the fluorescence anisotropy change on DNA binding indicated that apaMyoD bound to E-box-containing DNA sequences with enhanced affinity relative to MyoD-bHLH. Consequently, the DNA binding specificity of apaMyoD was increased 10-fold.

Original language	English
Pages (from-to)	69-77
Number of pages	9
Journal	Chemistry & Biology
Volume	11
Issue number	1
DOIs	https://doi.org/10.1016/j.chembiol.2003.12.015
Publication status	Published - Jan 2004

Keywords

Amino Acid Sequence
Anisotropy
Basic Helix-Loop-Helix Transcription Factors
Binding Sites
Circular Dichroism
DNA
DNA-Binding Proteins
Magnetic Resonance Spectroscopy
Microscopy, Fluorescence
Models, Molecular
Molecular Sequence Data
Protein Binding
Structure-Activity Relationship
Substrate Specificity
Temperature
Transcription Factors

Access to Document

10.1016/j.chembiol.2003.12.015

Cite this

@article{7092b4ad10244e2f972856c5c662cee6,

title = "Controlling the DNA binding specificity of bHLH proteins through intramolecular interactions",

abstract = "Reversible control of the conformation of proteins was employed to probe the relationship between flexibility and specificity of the basic helix-loop-helix protein MyoD. A fusion protein (apaMyoD) was designed where the basic DNA binding helix of MyoD was stablized by an amino-terminal extension with a sequence derived from the bee venom peptide apamin. The disulfide-stabilized helix from apamin served as a nucleus for a helix that extended for a further ten residues, thereby holding apaMyoD's DNA recognition helix in a predominantly alpha-helical conformation. The thermal stability of the DNA complexes of apaMyoD was increased by 13 degrees C relative to MyoD-bHLH. Measurements of the fluorescence anisotropy change on DNA binding indicated that apaMyoD bound to E-box-containing DNA sequences with enhanced affinity relative to MyoD-bHLH. Consequently, the DNA binding specificity of apaMyoD was increased 10-fold.",

keywords = "Amino Acid Sequence, Anisotropy, Basic Helix-Loop-Helix Transcription Factors, Binding Sites, Circular Dichroism, DNA, DNA-Binding Proteins, Magnetic Resonance Spectroscopy, Microscopy, Fluorescence, Models, Molecular, Molecular Sequence Data, Protein Binding, Structure-Activity Relationship, Substrate Specificity, Temperature, Transcription Factors",

author = "Turner, {Elizebeth C} and Cureton, {Charlotte H} and Weston, {Chris J} and Smart, {Oliver S} and Allemann, {Rudolf K}",

year = "2004",

month = jan,

doi = "10.1016/j.chembiol.2003.12.015",

language = "English",

volume = "11",

pages = "69--77",

journal = "Chemistry & Biology",

issn = "1074-5521",

publisher = "Elsevier",

number = "1",

}

TY - JOUR

T1 - Controlling the DNA binding specificity of bHLH proteins through intramolecular interactions

AU - Turner, Elizebeth C

AU - Cureton, Charlotte H

AU - Weston, Chris J

AU - Smart, Oliver S

AU - Allemann, Rudolf K

PY - 2004/1

Y1 - 2004/1

N2 - Reversible control of the conformation of proteins was employed to probe the relationship between flexibility and specificity of the basic helix-loop-helix protein MyoD. A fusion protein (apaMyoD) was designed where the basic DNA binding helix of MyoD was stablized by an amino-terminal extension with a sequence derived from the bee venom peptide apamin. The disulfide-stabilized helix from apamin served as a nucleus for a helix that extended for a further ten residues, thereby holding apaMyoD's DNA recognition helix in a predominantly alpha-helical conformation. The thermal stability of the DNA complexes of apaMyoD was increased by 13 degrees C relative to MyoD-bHLH. Measurements of the fluorescence anisotropy change on DNA binding indicated that apaMyoD bound to E-box-containing DNA sequences with enhanced affinity relative to MyoD-bHLH. Consequently, the DNA binding specificity of apaMyoD was increased 10-fold.

AB - Reversible control of the conformation of proteins was employed to probe the relationship between flexibility and specificity of the basic helix-loop-helix protein MyoD. A fusion protein (apaMyoD) was designed where the basic DNA binding helix of MyoD was stablized by an amino-terminal extension with a sequence derived from the bee venom peptide apamin. The disulfide-stabilized helix from apamin served as a nucleus for a helix that extended for a further ten residues, thereby holding apaMyoD's DNA recognition helix in a predominantly alpha-helical conformation. The thermal stability of the DNA complexes of apaMyoD was increased by 13 degrees C relative to MyoD-bHLH. Measurements of the fluorescence anisotropy change on DNA binding indicated that apaMyoD bound to E-box-containing DNA sequences with enhanced affinity relative to MyoD-bHLH. Consequently, the DNA binding specificity of apaMyoD was increased 10-fold.

KW - Amino Acid Sequence

KW - Anisotropy

KW - Basic Helix-Loop-Helix Transcription Factors

KW - Binding Sites

KW - Circular Dichroism

KW - DNA

KW - DNA-Binding Proteins

KW - Magnetic Resonance Spectroscopy

KW - Microscopy, Fluorescence

KW - Models, Molecular

KW - Molecular Sequence Data

KW - Protein Binding

KW - Structure-Activity Relationship

KW - Substrate Specificity

KW - Temperature

KW - Transcription Factors

U2 - 10.1016/j.chembiol.2003.12.015

DO - 10.1016/j.chembiol.2003.12.015

M3 - Article

C2 - 15112996

SN - 1074-5521

VL - 11

SP - 69

EP - 77

JO - Chemistry & Biology

JF - Chemistry & Biology

IS - 1

ER -

Controlling the DNA binding specificity of bHLH proteins through intramolecular interactions

Abstract

Keywords

Access to Document

Fingerprint

Cite this