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Nakamura, T. et al.

Multihormonal regulation of transcription of the tryptophan 2,3-dioxygenase gene in primary cultures of adult rat hepatocytes with special reference to the presence of a transcriptional protein mediating the action of glucocorticoids.

For study of hormonal regulation of gene expression of tryptophan 2,3-dioxygenase (EC 1. 13. 11. 11, TO), a DNA clone containing a sequence complementary to TO mRNA was prepared with TO mRNA from rat liver enriched 62-fold by immunoadsorption. Primary cultures of adult rat hepatocytes were treated with dexamethasone, and the amount of TO mRNA was measured by RNA dot-blot hybridization with this TO cDNA. Dexamethasone induced this TO mRNA 7-fold, while their treatments with dexamethasone plus glucagon induced the TO mRNA 18-fold. This induction of TO mRNA by dexamethasone plus glucagon was inhibited by insulin or epinephrine. Studies on transcription in isolated nuclei showed that these hormonal changes in the level of TO mRNA were caused by changes in the rate of transcription of the TO gene. Thus, expression of TO in the liver is regulated multihormonally at the transcriptional step. There was a long lag period before stimulation of transcription of the TO gene by dexamethasone in hepatocytes cultured for 20 h: the maximal rate was attained after 6-8 h. The lag time depended on the culture time without dexamethasone and was shorter after shorter culture of the cells. This finding suggested that a transcriptional factor that was lost during culture mediated the action of glucocorticoids. Consistent with this idea, cycloheximide or puromycin almost completely blocked enhanced transcription of the TO gene by dexamethasone after a 20-h culture, but not after a 2-h culture. These findings indicate that a short-lived transcriptional protein, which is also regulated by glucocorticoids, mediates their effect on expression of the TO gene.[1]

References

Multihormonal regulation of transcription of the tryptophan 2,3-dioxygenase gene in primary cultures of adult rat hepatocytes with special reference to the presence of a transcriptional protein mediating the action of glucocorticoids. Nakamura, T., Niimi, S., Nawa, K., Noda, C., Ichihara, A., Takagi, Y., Anai, M., Sakaki, Y. J. Biol. Chem. (1987) [Pubmed]

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