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PMID: 7651430 Published · ppublish English Comparative Study Journal Article

Exclusive homodimerization of the orphan receptor hepatocyte nuclear factor 4 defines a new subclass of nuclear receptors.

Molecular and cellular biology ·Vol. 15 ·No. 9 ·1995-09-00 ·Pages 5131-43

Jiang G, Nepomuceno L, Hopkins K, Sladek FM

Abstract

Hepatocyte nuclear factor 4 (HNF-4), a highly conserved member of the steroid hormone receptor superfamily critical for development and liver-specific gene expression, is very similar to another superfamily member, retinoid X receptor alpha (RXR alpha), in overall amino acid sequence and DNA binding specificity. Since RXR alpha is known to heterodimerize with many other nuclear receptors, the formation of heterodimers between HNF-4 and RXR alpha was examined. With the electrophoretic mobility shift assay, coimmunoprecipitation, and transient transfection assays, it is shown that, unlike other nuclear receptors, HNF-4 does not form heterodimers with RXR alpha either in the presence or in the absence of DNA. We also show that in vitro-translated HNF-4 does not form heterodimeric complexes on DNA with a number of other receptors, including RXR beta, RXR gamma, retinoic acid receptor alpha, or thyroid hormone receptor alpha. To investigate the hypothesis that the lack of heterodimerization between HNF-4 and RXR alpha is due to a strong homodimerization activity of HNF-4, glycerol gradient sedimentation and kinetic analysis were used to show that HNF-4 is in fact a stable homodimer in solution. Finally, immunohistochemistry is used to show that the HNF-4 protein is found exclusively in the nuclei in both HepG2 cells, which express endogenous HNF-4, and transfected COS cells, which overexpress HNF-4. These findings lead us to propose that HNF-4 defines a new subclass of nuclear receptors which reside primarily in the nucleus and which bind DNA and regulate transcription as homodimers.

MeSH Terms
Animals Base Sequence Cell Compartmentation Cells, Cultured DNA-Binding Proteins Hepatocyte Nuclear Factor 4 Immunohistochemistry Models, Biological Molecular Sequence Data Nuclear Proteins Peptide Fragments/metabolism Phosphoproteins Protein Binding Protein Conformation Receptors, Cytoplasmic and Nuclear/classification Receptors, Glucocorticoid/isolation & purification,metabolism Receptors, Retinoic Acid/metabolism Regulatory Sequences, Nucleic Acid Retinoid X Receptors Solutions Transcription Factors/isolation & purification,metabolism Transcription, Genetic
Chemicals
DNA-Binding Proteins Hepatocyte Nuclear Factor 4 Nuclear Proteins Peptide Fragments Phosphoproteins Receptors, Cytoplasmic and Nuclear Receptors, Glucocorticoid Receptors, Retinoic Acid Retinoid X Receptors Solutions Transcription Factors
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Jiang G
Environmental Toxicology Graduate Program, University of California, Riverside 92521, USA.
Nepomuceno L
Hopkins K
Sladek F M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.15.9.5131
Published
1995-09-00
Pages
5131-43
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC230760
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.15.9.5131 PMC Full Text View journal details
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