Interaction of triiodothyronine-receptor complexes with simian virus 40 minichromosomes in monkey kidney CV-1 cells.
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/micrograms SV40 minichromosomal DNA vs. 0.43 fmol/micrograms 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.
Savouret
J F
JF
Eberhardt
N L
NL
Cathala
G
G
Baxter
J D
JD
D000818:Animals; D001667:Binding, Competitive; D002467:Cell Nucleus / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002522:Chlorocebus aethiops; D002843:Chromatin / Q000378:metabolism; D002875:Chromosomes / Q000378:metabolism*; D003432:Cross-Linking Reagents; D005814:Genes, Viral; D013006:Growth Hormone / Q000235:genetics; D007668:Kidney / Q000382:microbiology* / Q000473:pathology; D012313:RNA / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011988:Receptors, Thyroid Hormone; D011995:Recombination, Genetic; D013539:Simian virus 40 / Q000235:genetics*; D014284:Triiodothyronine / Q000378:metabolism; D014412:Tumor Virus Infections / Q000473:pathology
D002843:Chromatin; D003432:Cross-Linking Reagents; D011956:Receptors, Cell Surface; D011988:Receptors, Thyroid Hormone; D014284:Triiodothyronine; D012313:RNA; D013006:Growth Hormone