Protein-Protein Interaction of Retinoic Acid Receptor α and Thyroid Transcription Factor-1 in Respiratory Epithelial Cells

Cong Yan, Angela Naltner, Julie Conkright, Manely Ghaffari

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Surfactant protein B (SP-B) is a 79-amino acid peptide critical to postnatal respiratory adaptation and is developmentally regulated. Previous studies demonstrated that retinoic acid receptors (RARs) and thyroid transcription factor 1 (TTF-1) stimulated SP-B gene expression in respiratory epithelial cells. Clustered retinoic acid-responsive element and TTF-1 binding sites were identified in the enhancer region of the SP-B gene and were required for retinoic acid stimulation of the human SP-B (hSP-B) promoter. In addition, RAR and TTF-1 were colocalized in mouse bronchiolar and alveolar type II epithelial cells, the cellular site of SP-B synthesis. In the present studies, RAR and TTF-1 were colocalized in the nucleus of H441 cells. RAR and TTF-1 synergistically stimulated the hSP-B promoter in H441 cells. Direct protein-protein interactions between RAR and TTF-1 were demonstrated by the glutathione S-transferase pull-down assay and the mammalian cell two hybrid assay. Truncation/deletion studies showed that the RAR-TTF-1 interaction was mediated through the RAR DNA binding domain (DBD) and the TTF-1 homeodomain. RAR DBD greatly enhanced TTF-1 homeodomain DNA binding activity to a hSP-B enhancer oligonucleotide, in which retinoic acid-responsive element and TTF-1 DNA binding sites overlap. Chromatin immunoprecipitation assay demonstrated that retinoic acid treatment of H441 cells greatly stimulated both RAR and TTF-1 DNA binding to the hSP-B enhancer region in H441 cells. These findings support a model in which RAR/retinoid X receptor, TTF-1, and coactivators (p160 members and CBP) form an enhanceosome in the enhancer region of the hSP-B gene.

Original languageEnglish (US)
Pages (from-to)21686-21691
Number of pages6
JournalJournal of Biological Chemistry
Volume276
Issue number24
StatePublished - Jun 15 2001
Externally publishedYes

Fingerprint

Retinoic Acid Receptors
Epithelial Cells
Proteins
Surface-Active Agents
Tretinoin
DNA
Assays
thyroid nuclear factor 1
Genes
Binding Sites
Retinoid X Receptors
Alveolar Epithelial Cells
Two-Hybrid System Techniques
Chromatin Immunoprecipitation
Glutathione Transferase
Cell Nucleus
Gene expression
Oligonucleotides
Chromatin
Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Protein-Protein Interaction of Retinoic Acid Receptor α and Thyroid Transcription Factor-1 in Respiratory Epithelial Cells. / Yan, Cong; Naltner, Angela; Conkright, Julie; Ghaffari, Manely.

In: Journal of Biological Chemistry, Vol. 276, No. 24, 15.06.2001, p. 21686-21691.

Research output: Contribution to journalArticle

Yan, Cong ; Naltner, Angela ; Conkright, Julie ; Ghaffari, Manely. / Protein-Protein Interaction of Retinoic Acid Receptor α and Thyroid Transcription Factor-1 in Respiratory Epithelial Cells. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 24. pp. 21686-21691.
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