This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via Google Scholar Google Scholar Articles by SAVOURET, J.-F. Articles by BAXTER, J. D. Search for Related Content PubMed PubMed Citation Articles by SAVOURET, J.-F. Articles by BAXTER, J. D.

Interaction of Triiodothyronine-Receptor Complexes with Simian Virus 40 Minichromosomes in Monkey Kidney CV-1 Cells^*

JEAN-FRANCOIS SAVOURET, NORMAN L. EBERHARDT, GUY CATHALA and JOHN D. BAXTER

Metabolic Research Unit University of California San Francisco, California 94143
Departments of Medicine University of California San Francisco, California 94143
Biochemistry and Biophysics University of California San Francisco, California 94143

Address all correspondence and requests for reprints to: Norman L. Eberhardt, Metabolic Research Unit, 1141-HSW, University of California, San Francisco, California 94143.

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 SV40 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/Hg 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. (Endocrinology 116: 1259-1266,1985)

Footnotes

^* This work was supported in part by NIH Grants AM-18878, AM-19997, and HD-17838.

Supported by a fellowship from the Delegation Generate a la Recherche Scientifique et Technique. Present address: INSERM Unit 135, Hopital de Bicetre, 78 Avenue du General Leclerc, 94270 Kremlin-Bicetre, France.

Present address: Laboratoire de Biologie Moleculaire, USTL, Place Eugene Bataillon, 34060 Montpellier Cedex, France.

Received August 17, 1984.