Influence of domain interactions on conformational mobility of the progesterone receptor detected by hydrogen/deuterium exchange mass spectrometry

Structure. 2014 Jul 8;22(7):961-73. doi: 10.1016/j.str.2014.04.013. Epub 2014 Jun 5.

Abstract

Structural and functional details of the N-terminal activation function 1 (AF1) of most nuclear receptors are poorly understood due to the highly dynamic intrinsically disordered nature of this domain. A hydrogen/deuterium exchange (HDX) mass-spectrometry-based investigation of TATA box-binding protein (TBP) interaction with various domains of progesterone receptor (PR) demonstrate that agonist-bound PR interaction with TBP via AF1 impacts the mobility of the C-terminal AF2. Results from HDX and other biophysical studies involving agonist- and antagonist-bound full-length PR and isolated PR domains reveal the molecular mechanism underlying synergistic transcriptional activation mediated by AF1 and AF2, dominance of PR-B isoform over PR-A, and the necessity of AF2 for full AF1-mediated transcriptional activity. These results provide a comprehensive picture elaborating the underlying mechanism of PR-TBP interactions as a model for studying nuclear receptor (NR)-transcription factor functional interactions.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Deuterium Exchange Measurement
  • Humans
  • Ligands
  • Mass Spectrometry / methods*
  • Mifepristone / chemistry
  • Mifepristone / metabolism
  • Models, Molecular
  • Molecular Sequence Data
  • Promegestone / chemistry
  • Promegestone / metabolism
  • Protein Binding
  • Protein Conformation*
  • Protein Structure, Tertiary*
  • Receptors, Cytoplasmic and Nuclear / chemistry
  • Receptors, Cytoplasmic and Nuclear / metabolism
  • Receptors, Progesterone / chemistry*
  • Receptors, Progesterone / genetics
  • Receptors, Progesterone / metabolism
  • Sf9 Cells
  • TATA-Box Binding Protein / chemistry
  • TATA-Box Binding Protein / metabolism

Substances

  • Ligands
  • Receptors, Cytoplasmic and Nuclear
  • Receptors, Progesterone
  • TATA-Box Binding Protein
  • TBP protein, human
  • Mifepristone
  • Promegestone