Conformational analysis of the androgen receptor amino-terminal domain involved in transactivation. Influence of structure-stabilizing solutes and protein-protein interactions

Jill Reid, S. M. Kelly, K. Watt, N. C. Price, Iain Joseph McEwan

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93 Citations (Scopus)

Abstract

The androgen receptor (AR) is a member of the nuclear receptor superfamily. Sequences within the large amino-terminal domain of the receptor have been shown to be important for transactivation and protein-protein interactions; however, little is known about the structure and folding of this region. In the present study we show that a 344-amino acid polypeptide representing the main determinants for transactivation has the propensity to form a-helical structure and that mutations which disrupt putative helical regions alter conformation. Folding of the AR was observed in the presence of the helix-stabilizing solvent trifluoroethanol and the natural osmolyte trimethylamine N-oxide (TMAO). TMAO resulted in the movement of two tryptophan residues to a less solvent-exposed environment and the formation of secondary/tertiary structure resistant to protease cleavage. Critically, binding to the RAP74 subunit of the general transcription factor TFIIF resulted in extensive protease resistance, consistent with induced folding of the receptor transactivation domain. These data indicate that this region of the AR is structurally flexible and folds into a stable conformation upon interactions with a component of the general transcription machinery.

Original languageEnglish
Pages (from-to)20079-20086
Number of pages7
JournalThe Journal of Biological Chemistry
Volume277
Issue number22
DOIs
Publication statusPublished - May 2002

Keywords

  • TRANSCRIPTIONAL ACTIVATION DOMAIN
  • HUMAN GLUCOCORTICOID RECEPTOR
  • LIGAND-BINDING DOMAIN
  • NUCLEAR RECEPTOR
  • SECONDARY STRUCTURE
  • ALPHA-HELIX
  • IN-VITRO
  • COACTIVATOR
  • REGIONS
  • VP16

Cite this

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title = "Conformational analysis of the androgen receptor amino-terminal domain involved in transactivation. Influence of structure-stabilizing solutes and protein-protein interactions",
abstract = "The androgen receptor (AR) is a member of the nuclear receptor superfamily. Sequences within the large amino-terminal domain of the receptor have been shown to be important for transactivation and protein-protein interactions; however, little is known about the structure and folding of this region. In the present study we show that a 344-amino acid polypeptide representing the main determinants for transactivation has the propensity to form a-helical structure and that mutations which disrupt putative helical regions alter conformation. Folding of the AR was observed in the presence of the helix-stabilizing solvent trifluoroethanol and the natural osmolyte trimethylamine N-oxide (TMAO). TMAO resulted in the movement of two tryptophan residues to a less solvent-exposed environment and the formation of secondary/tertiary structure resistant to protease cleavage. Critically, binding to the RAP74 subunit of the general transcription factor TFIIF resulted in extensive protease resistance, consistent with induced folding of the receptor transactivation domain. These data indicate that this region of the AR is structurally flexible and folds into a stable conformation upon interactions with a component of the general transcription machinery.",
keywords = "TRANSCRIPTIONAL ACTIVATION DOMAIN, HUMAN GLUCOCORTICOID RECEPTOR, LIGAND-BINDING DOMAIN, NUCLEAR RECEPTOR, SECONDARY STRUCTURE, ALPHA-HELIX, IN-VITRO, COACTIVATOR, REGIONS, VP16",
author = "Jill Reid and Kelly, {S. M.} and K. Watt and Price, {N. C.} and McEwan, {Iain Joseph}",
year = "2002",
month = "5",
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language = "English",
volume = "277",
pages = "20079--20086",
journal = "The Journal of Biological Chemistry",
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T1 - Conformational analysis of the androgen receptor amino-terminal domain involved in transactivation. Influence of structure-stabilizing solutes and protein-protein interactions

AU - Reid, Jill

AU - Kelly, S. M.

AU - Watt, K.

AU - Price, N. C.

AU - McEwan, Iain Joseph

PY - 2002/5

Y1 - 2002/5

N2 - The androgen receptor (AR) is a member of the nuclear receptor superfamily. Sequences within the large amino-terminal domain of the receptor have been shown to be important for transactivation and protein-protein interactions; however, little is known about the structure and folding of this region. In the present study we show that a 344-amino acid polypeptide representing the main determinants for transactivation has the propensity to form a-helical structure and that mutations which disrupt putative helical regions alter conformation. Folding of the AR was observed in the presence of the helix-stabilizing solvent trifluoroethanol and the natural osmolyte trimethylamine N-oxide (TMAO). TMAO resulted in the movement of two tryptophan residues to a less solvent-exposed environment and the formation of secondary/tertiary structure resistant to protease cleavage. Critically, binding to the RAP74 subunit of the general transcription factor TFIIF resulted in extensive protease resistance, consistent with induced folding of the receptor transactivation domain. These data indicate that this region of the AR is structurally flexible and folds into a stable conformation upon interactions with a component of the general transcription machinery.

AB - The androgen receptor (AR) is a member of the nuclear receptor superfamily. Sequences within the large amino-terminal domain of the receptor have been shown to be important for transactivation and protein-protein interactions; however, little is known about the structure and folding of this region. In the present study we show that a 344-amino acid polypeptide representing the main determinants for transactivation has the propensity to form a-helical structure and that mutations which disrupt putative helical regions alter conformation. Folding of the AR was observed in the presence of the helix-stabilizing solvent trifluoroethanol and the natural osmolyte trimethylamine N-oxide (TMAO). TMAO resulted in the movement of two tryptophan residues to a less solvent-exposed environment and the formation of secondary/tertiary structure resistant to protease cleavage. Critically, binding to the RAP74 subunit of the general transcription factor TFIIF resulted in extensive protease resistance, consistent with induced folding of the receptor transactivation domain. These data indicate that this region of the AR is structurally flexible and folds into a stable conformation upon interactions with a component of the general transcription machinery.

KW - TRANSCRIPTIONAL ACTIVATION DOMAIN

KW - HUMAN GLUCOCORTICOID RECEPTOR

KW - LIGAND-BINDING DOMAIN

KW - NUCLEAR RECEPTOR

KW - SECONDARY STRUCTURE

KW - ALPHA-HELIX

KW - IN-VITRO

KW - COACTIVATOR

KW - REGIONS

KW - VP16

U2 - 10.1074/jbc.M201003200

DO - 10.1074/jbc.M201003200

M3 - Article

VL - 277

SP - 20079

EP - 20086

JO - The Journal of Biological Chemistry

JF - The Journal of Biological Chemistry

SN - 0021-9258

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ER -