Analysis of protein dimerization and ligand binding of orphan receptor HNF4α

Andrew A. Bogan, Qing Dallas-Yang, Michael D. Ruse, Yutaka Maeda, Guoqiang Jiang, Luviminda Nepomuceno, Thomas (Tom) Scanlan, Fred E. Cohen, Frances M. Sladek

Research output: Contribution to journalArticle

89 Citations (Scopus)

Abstract

Hepatocyte nuclear factor 4α (HNF4α) (NR2A1), an orphan member of the nuclear receptor superfamily, binds DNA exclusively as a homodimer even though it is very similar in amino acid sequence to retinoid X receptor α (RXRα), which heterodimerizes readily with other receptors. Here, experimental analysis of residues involved in protein dimerization and studies on a reported ligand for HNF4α are combined with a structural model of the HNF4α ligand-binding domain (LBD) (residues 137 to 384). When K300 (in helix 9) and E327 (in helix 10) of HNF4α1 were converted to the analogous residues in-RXRα (E390 and K417, respectively) the resulting construct did not heterodimerize with the wild-type HNF4α, although it was still able to form homodimers and bind DNA. Furthermore, the double mutant did not heterodimerize with RXR or RAR but was still able to dimerize in solution with an HNF4α construct truncated at amino acid residue 268. This suggests that the charge compatibility between helices 9 and 10 is necessary, but not sufficient, to determine dimerization partners, and that additional residues in the HNF4α LBD are also important in dimerization. The structural model of the HNF4α LBD and an amino acid sequence alignment of helices 9 and 10 in various HNF4 and other receptor genes indicates that a K(X)26E motif can be used to identify HNF4 genes from other organisms and that a (E/D(X)26-29K/R) motif can be used to predict heterodimerization of many, but not all, receptors with RXR. In vitro analysis of another HNF4α mutant construct indicates that helix 10 also plays a structural role in the conformational integrity of HNF4α. The structural model and experimental analysis indicate that fatty acyl CoA thioesters, the proposed HNF4α ligands, are not good candidates for a traditional ligand for HNF4α. Finally, these results provide insight into the mechanism of action of naturally occurring mutations in the human HNF4α gene found in patients with maturity onset diabetes of the young 1 (MODY1). (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)831-851
Number of pages21
JournalJournal of Molecular Biology
Volume302
Issue number4
DOIs
StatePublished - Sep 29 2000
Externally publishedYes

Fingerprint

Hepatocyte Nuclear Factor 4
Protein Multimerization
Ligands
Retinoid X Receptors
Structural Models
Dimerization
Amino Acid Sequence
Orphan Nuclear Receptors
Genes
Acyl Coenzyme A
Sequence Alignment
DNA

Keywords

  • HNF4
  • Ligand
  • MODY1
  • Receptor dimerization

ASJC Scopus subject areas

  • Virology

Cite this

Bogan, A. A., Dallas-Yang, Q., Ruse, M. D., Maeda, Y., Jiang, G., Nepomuceno, L., ... Sladek, F. M. (2000). Analysis of protein dimerization and ligand binding of orphan receptor HNF4α. Journal of Molecular Biology, 302(4), 831-851. https://doi.org/10.1006/jmbi.2000.4099

Analysis of protein dimerization and ligand binding of orphan receptor HNF4α. / Bogan, Andrew A.; Dallas-Yang, Qing; Ruse, Michael D.; Maeda, Yutaka; Jiang, Guoqiang; Nepomuceno, Luviminda; Scanlan, Thomas (Tom); Cohen, Fred E.; Sladek, Frances M.

In: Journal of Molecular Biology, Vol. 302, No. 4, 29.09.2000, p. 831-851.

Research output: Contribution to journalArticle

Bogan, AA, Dallas-Yang, Q, Ruse, MD, Maeda, Y, Jiang, G, Nepomuceno, L, Scanlan, TT, Cohen, FE & Sladek, FM 2000, 'Analysis of protein dimerization and ligand binding of orphan receptor HNF4α', Journal of Molecular Biology, vol. 302, no. 4, pp. 831-851. https://doi.org/10.1006/jmbi.2000.4099
Bogan AA, Dallas-Yang Q, Ruse MD, Maeda Y, Jiang G, Nepomuceno L et al. Analysis of protein dimerization and ligand binding of orphan receptor HNF4α. Journal of Molecular Biology. 2000 Sep 29;302(4):831-851. https://doi.org/10.1006/jmbi.2000.4099
Bogan, Andrew A. ; Dallas-Yang, Qing ; Ruse, Michael D. ; Maeda, Yutaka ; Jiang, Guoqiang ; Nepomuceno, Luviminda ; Scanlan, Thomas (Tom) ; Cohen, Fred E. ; Sladek, Frances M. / Analysis of protein dimerization and ligand binding of orphan receptor HNF4α. In: Journal of Molecular Biology. 2000 ; Vol. 302, No. 4. pp. 831-851.
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