Post-transcriptional inactivation of p53 in immortalized murine embryo fibroblast cells

Hyunggee Kim, Seungkwon You, James Farris, Linda K. Foster, Douglas N. Foster

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The steady-state levels of p53 mRNA and protein were barely detectable by Northern and Western blot analysis in spontaneously immortalized (10)3 and (10)7 murine embryo fibroblast (MEF) cells. But when cells were treated with cycloheximide (CHX) or emetine, expression levels were restored to those observed in primary and immortal (10)10 MEF cells. However, levels of p53 mRNA were not changed in primary or (10)10 MEF cells by CHX treatment. De novo p53 mRNA synthetic rates were similar in primary, (10)10, (10)3, and (10)7 MEF cells treated with or without CHX. Treatment with actinomycin D (ActD) showed that p53 mRNA in primary and (10)10 MEF cells had a relatively long half-life of 22 h, compared to less than 2 h for (10)3 and (10)7 MEF cells. Pulse-chase analysis of p53 mRNA turnover using CHX and ActD showed that the rapid destabilization of p53 mRNA in (10)3 and (10)7 MEF cells could be regulated at the transcriptional and translational levels. In addition, the destabilization of p53 mRNA appeared to occur in the nucleus for (10)3 and (10)7 cells, but not for primary and (10)10 MEF cells. Taken together, the present study demonstrates that inactivation of the p53 gene occurs at the post-transcriptional level by rapid destabilization of its mRNA in the nucleus of spontaneously immortalized (10)3 and (10)7 MEF cells.

Original languageEnglish
Pages (from-to)3306-3310
Number of pages5
JournalOncogene
Volume20
Issue number25
DOIs
Publication statusPublished - 2001
Externally publishedYes

Keywords

  • Actinomycin D
  • Cycloheximide
  • Emetine
  • Half-life
  • Immortal MEF
  • P53 mRNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

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