Cell cycle-dependent expression of thyroid hormone receptor-beta is a mechanism for variable hormone sensitivity.
Thyroid hormone receptors (TRs) are ligand-regulatable transcription factors. Currently, little is known about the expression of TRs or other nuclear hormone receptors during the cell cycle. We thus developed a stable expression system to express green fluorescent protein-TRbeta in HeLa cells under tetracycline regulation, and studied TR expression during the cell cycle by laser scanning cytometry. Only approximately 9-15% of the nonsynchronized cell population expressed TR because the majority of cells were in G(1) phase and did not express detectable amounts of TR. However, when cells were synchronized in early S phase with hydroxyurea and then released, TR expression levels increased in a cell cycle-dependent manner and peaked to 30-40% cells expressing TR at late G(2)/M phase before declining to nonsynchronized levels. Moreover, we observed a direct correlation between transcriptional activity and TR expression during the cell cycle. Similar cell cycle-dependent findings also were observed for endogenous TR in rat pituitary GH(3) cells. Last, cycloheximide studies demonstrated that the increase in TR expression was primarily due to increased translation. These novel observations of cell cycle-dependent expression of TR suggest that differential hormone sensitivity can occur during the cell cycle and may contribute to cell cycle progression during normal development and oncogenesis.
Maruvada
Padma
P
Molecular Regulation and Neuroendocrinology Section, Clinical Endocrinology Branch, National Institute of Diabetes, Digestive, and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.
Dmitrieva
Natalia I
NI
East-Palmer
Joyce
J
Yen
Paul M
PM
D000818:Animals; D015153:Blotting, Western; D002453:Cell Cycle; D002467:Cell Nucleus / Q000378:metabolism; D002469:Cell Separation; D003513:Cycloheximide / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004318:Doxycycline / Q000494:pharmacology; D005434:Flow Cytometry; D016193:G1 Phase; D016195:G2 Phase; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006728:Hormones / Q000378:metabolism; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology; D007834:Lasers; D008024:Ligands; D008164:Luminescent Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008938:Mitosis; D010957:Plasmids / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D051381:Rats; D011988:Receptors, Thyroid Hormone / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016196:S Phase; D013752:Tetracycline / Q000494:pharmacology; D037042:Thyroid Hormone Receptors beta; D013997:Time Factors; D014158:Transcription, Genetic; D015533:Transcriptional Activation
D006728:Hormones; D008024:Ligands; D008164:Luminescent Proteins; D011988:Receptors, Thyroid Hormone; D037042:Thyroid Hormone Receptors beta; D049452:Green Fluorescent Proteins; D003513:Cycloheximide; D013752:Tetracycline; D004318:Doxycycline; D006918:Hydroxyurea