Van Heuverswyn B, Streydio C, Brocas H, Refetoff S, Dumont J, Vassart G
Proc Natl Acad Sci U S A. 1984 Oct;81(19):5941-5. doi: 10.1073/pnas.81.19.5941.
The availability of rat thyroglobulin cDNA clones was exploited to study the regulation of thyroglobulin gene transcription by thyrotropin (TSH). Groups of rats were subjected to treatments leading to reduction or increase in the rat serum TSH (rTSH) levels. Thyroid gland nuclei were isolated, incubated in vitro in the presence of 32P-labeled uridine triphosphate, and thyroglobulin transcripts were quantitated by hybridization to immobilized rat thyroglobulin cDNA clones. Transcription of the thyroglobulin gene was found to be very active in thyroid nuclei from control animals. It represented about 10% of total RNA polymerase II activity. Chronic hyperstimulation of the thyroid glands with endogenous rTSH was achieved in rats treated with the goitrogen propylthiouracil. No significant increase of thyroglobulin gene transcription could be measured in thyroid nuclei from these animals. On the contrary, a dramatic decrease in thyroglobulin gene transcription was observed in those animals in which endogenous rTSH levels had been suppressed by hypophysectomy or by the administration of triiodothyronine. Injection of exogenous bovine TSH in such animals readily restored transcriptional activity of the gene. Our results identify transcription as an important regulatory step involved in TSH action. They suggest that normal TSH levels induce close to maximal expression of the thyroglobulin gene but that continuous presence of TSH is required in order to maintain the gene in an activated state.
利用大鼠甲状腺球蛋白cDNA克隆来研究促甲状腺激素(TSH)对甲状腺球蛋白基因转录的调控。将大鼠分组进行处理,以降低或升高大鼠血清TSH(rTSH)水平。分离甲状腺腺细胞核,在含有32P标记的尿苷三磷酸的条件下进行体外培养,通过与固定化的大鼠甲状腺球蛋白cDNA克隆杂交来定量甲状腺球蛋白转录本。发现甲状腺球蛋白基因在对照动物的甲状腺细胞核中转录非常活跃。它约占总RNA聚合酶II活性的10%。在用致甲状腺肿物质丙硫氧嘧啶处理的大鼠中,实现了内源性rTSH对甲状腺的慢性过度刺激。在这些动物的甲状腺细胞核中未检测到甲状腺球蛋白基因转录的显著增加。相反,在那些通过垂体切除或给予三碘甲状腺原氨酸抑制了内源性rTSH水平的动物中,观察到甲状腺球蛋白基因转录显著下降。在此类动物中注射外源性牛TSH可迅速恢复该基因的转录活性。我们的结果确定转录是TSH作用中涉及的一个重要调控步骤。它们表明正常的TSH水平可诱导甲状腺球蛋白基因接近最大程度的表达,但为了使该基因维持在激活状态,需要持续存在TSH。