Hormonal regulation of transcription of rDNA. Inhibtion of transcription during glucocorticoid-mediated inhibition of proliferation of lymphosarcoma P1798 cells in culture

A. H. Cavanaugh, E. A. Thompson

Research output: Contribution to journalArticle

30 Scopus citations

Abstract

A lymphosarcoma cell line designated P1798.S6M has been characterized with respect to glucocorticod responsiveness in culture. Cells ceased to proliferate in the presence of 10-7 M dexamethasone. Cell viability remained high and glucocorticoid-sensitive cells could be rescued from cultures treated with dexamethasone for 24 or 48 h. These data indicate that P1798.S6M undergoes reversible arrest in the presence of dexamethasone. The system was used to study the effects of mitotic arrest upon transcription of rDNA. Incorporaton of [methyl-3H]methionine into rRNA was rapidly inhibited and pulse-chase experiments indicated that 28 S RNA was not synthesized after 24 h of exposure to dexamethasone. Hybridization studies indicated that the amount of pre-rRNA was reduced by 90 to 95% in cells treated for 24 h. Transcription studies were carried out in isolated nuclei. Twenty-four hours after addition of dexamethasone, template-bound RNA polymerase I activity decreased by 89% to 96%. Total RNA polymerase I activity did not decrease, whereas disengaged nuclear polymerase I activity increased dramatically. Filter hybridization studies indicated that labeling of nascent pre-rRNA chains in vitro was inhibited 93%. These data are interpreted as follows: Dexamethasone reversibly inhibits proliferation of P1798.S6M cells and transcription of rDNA. Total RNA polymerase I activity does not decrease, but the amount of template-bound enzyme is reduced with a concomitant increase in the amount of disengaged polymerase I. This indicates that initiation of transcription is inhibited in cells undergoing mitotic arrest in the presence of dexamethasone.

Original languageEnglish (US)
Pages (from-to)9768-9773
Number of pages6
JournalJournal of Biological Chemistry
Volume258
Issue number16
StatePublished - Jan 1 1983

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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