Molecular basis of coiled coil coactivator recruitment by the aryl hydrocarbon receptor nuclear translocator (ARNT)

J Biol Chem. 2009 May 29;284(22):15184-92. doi: 10.1074/jbc.M808479200. Epub 2009 Mar 26.

Abstract

The aryl hydrocarbon receptor nuclear translocator (ARNT) serves as the obligate heterodimeric partner for bHLH-PAS proteins involved in sensing and coordinating transcriptional responses to xenobiotics, hypoxia, and developmental pathways. Although its C-terminal transactivation domain is dispensable for transcriptional activation in vivo, ARNT has recently been shown to use its N-terminal bHLH and/or PAS domains to interact with several transcriptional coactivators that are required for transcriptional initiation after xenobiotic or hypoxic cues. Here we show that ARNT uses a single PAS domain to interact with two coiled coil coactivators, TRIP230 and CoCoA. Both coactivators interact with the same interface on the ARNT PAS-B domain, located on the opposite side of the domain used to associate with the analogous PAS domain on its heterodimeric bHLH-PAS partner HIF-2alpha. Using NMR and biochemical studies, we identified the ARNT-interacting motif of one coactivator, TRIP230 as an LXXLL-like nuclear receptor box. Mutation of this motif and proximal sequences disrupts the interaction with ARNT PAS-B. Identification of this ARNT-coactivator interface illustrates how ARNT PAS-B is used to form critical interactions with both bHLH-PAS partners and coactivators that are required for transcriptional responses.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Aryl Hydrocarbon Receptor Nuclear Translocator / chemistry
  • Aryl Hydrocarbon Receptor Nuclear Translocator / metabolism*
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Binding Sites
  • Conserved Sequence
  • Humans
  • Magnetic Resonance Spectroscopy
  • Molecular Sequence Data
  • Mutation / genetics
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / metabolism*
  • Peptides / chemistry
  • Protein Binding
  • Protein Interaction Mapping
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Trans-Activators / chemistry
  • Trans-Activators / metabolism*

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • Nuclear Proteins
  • Peptides
  • Trans-Activators
  • Aryl Hydrocarbon Receptor Nuclear Translocator
  • endothelial PAS domain-containing protein 1