Interaction of triiodothyronine-receptor complexes with simian virus 40 minichromosomes in monkey kidney CV-1 cells

J. F. Savouret, N. L. Eberhardt, G. Cathala, J. D. Baxter

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Thyroid hormone-responsive tissues contain chromatin-localized receptors that bind to DNA and may associate preferentially with actively transcribed chromatin. To study such receptor-chromatin localization, we have used cultured CV-1 cells permissive for simian virus 40 (SV40), in which viral minichromosomes can be separated from the cellular chromatin. CV-1 cells were found to contain intranuclear thyroid hormone-binding sites with an affinity for T3 and T4 and a site concentration similar to those in other thyroid hormone-responsive tissues. When these cells were infected with SV40 or an SV40-human GH gene recombinant, T3 did not affect SV 40 replication, early or late gene transcription, or human GH gene expression. However, in both cases, these infections resulted in the association of about 7.5% of the total specific T3-binding activity with the SV40 minichromosome, representing about 1 receptor molecule/65 minichromosomes and a 10-fold enrichment over the cellular chromatin-associated activity (4.3 fmol/μg SV40 minichromosomal DNA vs. 0.43 fmol/μg chromosomal DNA); 30% of this could be covalently cross-linked to the minichromosome with dissuccinimidyl suberate. The minichromosomes were also found to be transcriptionally active. Thus, thyroid hormone receptors interact preferentially with the SV40 minichromosome, possibly owing to their tendency to associate with transcriptionally active chromatin. This system provides an alternate approach to study the association of thyroid hormone receptors with defined chromosomal segments.

Original languageEnglish (US)
Pages (from-to)1259-1266
Number of pages8
JournalEndocrinology
Volume116
Issue number4
StatePublished - 1985
Externally publishedYes

Fingerprint

Thyroid Hormone Receptors
Simian virus 40
Chromatin
Haplorhini
Kidney
Thyroid Hormones
Genetic Transcription
DNA
1,2-diamino-1,2-N,N'-carbonyl-1,2-dideoxyglucose hydrate
Binding Sites
Gene Expression
Infection
Genes

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Savouret, J. F., Eberhardt, N. L., Cathala, G., & Baxter, J. D. (1985). Interaction of triiodothyronine-receptor complexes with simian virus 40 minichromosomes in monkey kidney CV-1 cells. Endocrinology, 116(4), 1259-1266.

Interaction of triiodothyronine-receptor complexes with simian virus 40 minichromosomes in monkey kidney CV-1 cells. / Savouret, J. F.; Eberhardt, N. L.; Cathala, G.; Baxter, J. D.

In: Endocrinology, Vol. 116, No. 4, 1985, p. 1259-1266.

Research output: Contribution to journalArticle

Savouret, JF, Eberhardt, NL, Cathala, G & Baxter, JD 1985, 'Interaction of triiodothyronine-receptor complexes with simian virus 40 minichromosomes in monkey kidney CV-1 cells', Endocrinology, vol. 116, no. 4, pp. 1259-1266.
Savouret, J. F. ; Eberhardt, N. L. ; Cathala, G. ; Baxter, J. D. / Interaction of triiodothyronine-receptor complexes with simian virus 40 minichromosomes in monkey kidney CV-1 cells. In: Endocrinology. 1985 ; Vol. 116, No. 4. pp. 1259-1266.
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