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Yeast As a Chassis for Developing Functional Assays to Study Human P53
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P53 regulates CCAAT/Enhancer binding protein β gene expression.

Biao Hu1, Tianju Liu2, Zhe Wu2

  • 1Department of Internal Medicine, University of Michigan Medical School, 1600 Huron Parkway, Ann Arbor, MI 48109 USA.

Gene
|August 4, 2023
PubMed
Summary

The tumor suppressor P53 represses CCAAT/enhancer-binding protein β (C/EBPβ) gene expression. P53 controls both C/EBPβ transcription and translation, affecting its LAP and LIP isoforms differently.

Keywords:
CCAAT/Enhancer Binding Protein βP53Transcriptional regulationTranslational regulationeIF4e

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Area of Science:

  • Molecular Biology
  • Cancer Biology
  • Gene Regulation

Background:

  • CCAAT/enhancer-binding protein β (C/EBPβ) is a transcription factor involved in various cellular processes and diseases.
  • C/EBPβ exists in two isoforms, LAP and LIP, translated from the same mRNA, with LIP inhibiting LAP activity.
  • The regulation of C/EBPβ gene expression and the LAP:LIP ratio remains unclear.

Purpose of the Study:

  • To investigate the role of P53 in regulating C/EBPβ gene expression.
  • To elucidate the mechanism by which P53 affects C/EBPβ transcription and translation.
  • To determine if P53 differentially regulates C/EBPβ LAP and LIP isoforms.

Main Methods:

  • Promoter analysis to identify P53 response elements (P53RE) in the C/EBPβ gene.
  • Electrophoretic Mobility Shift Assay (EMSA) and chromatin immunoprecipitation to confirm P53 binding.
  • Gene transfection studies with P53 overexpression and dominant-negative P53.
  • Western blot analysis to assess C/EBPβ isoform expression.
  • 5' untranslated region (5'UTR) constructs to study translational control.

Main Results:

  • A conserved P53RE was identified in the C/EBPβ promoter, with P53 binding confirmed by EMSA and ChIP.
  • P53 overexpression suppressed C/EBPβ transcription and expression, while dominant-negative P53 enhanced it.
  • P53 differentially regulated the translation of LAP and LIP isoforms via eIF4E and eIF4E-BP1.
  • The 5'UTR of C/EBPβ plays a crucial role in the differential translation of LAP and LIP isoforms.

Conclusions:

  • P53 acts as a repressor of C/EBPβ gene expression at both transcriptional and translational levels.
  • P53 exhibits differential regulation of C/EBPβ LAP and LIP isoforms.
  • This study reveals a novel mechanism for P53 antagonizing C/EBPβ activity.