Isolation and characterization of a novel coactivator protein, NCoA-62, involved in vitamin D-mediated transcription

T. A. Baudino, D. M. Kraichely, Jr Jefcoat S.C., S. K. Winchester, Nicola Partridge, P. N. MacDonald

Research output: Contribution to journalArticle

Abstract

The vitamin D receptor (VDR) forms a heterodimeric complex with retinoid X receptor (RXR) and binds to vitamin D-responsive promoter elements to regulate the transcription of specific genes or gene networks. The precise mechanism of transcriptional regulation by the VDR·RXR heterodimer is not well understood, but it may involve interactions of VDR·RXR with transcriptional coactivator or corepressor proteins. Here, a yeast two- hybrid strategy was used to isolate proteins that selectively interacted with VDR and other nuclear receptors. One cDNA clone designated NCoA-62, encoded a 62,000-Da protein that is highly related to BX42, a Drosophila melanogaster nuclear protein involved in ecdysone-stimulated gene expression. Yeast two- hybrid studies and in vitro protein-protein interaction assays using glutathione S-transferase fusion proteins demonstrated that NCoA-62 formed a direct protein-protein contact with the ligand binding domain of VDR. Coexpression of NCoA-62 in a vitamin D-responsive transient gene expression system augmented 1,25-dihydroxy-vitamin D3-activated transcription, but it had little or no effect on basal transcription or gal4-VP16-activated transcription. NCoA-62 also interacted with retinoid receptors, and its expression enhanced retinoic acid-, estrogen-, and glucocorticoid-mediated gene expression. These data indicate that NCoA-62 may be classified into an emerging set of transcriptional coactivator proteins that function to facilitate vitamin D- and other nuclear receptor-mediated transcriptional pathways.

Original languageEnglish (US)
Pages (from-to)16434-16441
Number of pages8
JournalJournal of Biological Chemistry
Volume273
Issue number26
DOIs
StatePublished - Jun 26 1998

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Transcription
Vitamin D
Calcitriol Receptors
Proteins
Gene expression
Cytoplasmic and Nuclear Receptors
Gene Expression
Yeast
Genes
Yeasts
Retinoid X Receptors
Ecdysone
Drosophila Proteins
Co-Repressor Proteins
Gene Regulatory Networks
Retinoids
Nuclear Proteins
Tretinoin
Glutathione Transferase
Glucocorticoids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Isolation and characterization of a novel coactivator protein, NCoA-62, involved in vitamin D-mediated transcription. / Baudino, T. A.; Kraichely, D. M.; Jefcoat S.C., Jr; Winchester, S. K.; Partridge, Nicola; MacDonald, P. N.

In: Journal of Biological Chemistry, Vol. 273, No. 26, 26.06.1998, p. 16434-16441.

Research output: Contribution to journalArticle

Baudino, T. A. ; Kraichely, D. M. ; Jefcoat S.C., Jr ; Winchester, S. K. ; Partridge, Nicola ; MacDonald, P. N. / Isolation and characterization of a novel coactivator protein, NCoA-62, involved in vitamin D-mediated transcription. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 26. pp. 16434-16441.
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