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Related Experiment Videos

CREB controls LAP/C/EBP beta transcription

M Niehof1, M P Manns, C Trautwein

  • 1Department of Gastroenterology and Hepatology, Medizinische Hochschule Hannover, Germany.

Molecular and Cellular Biology
|July 1, 1997
PubMed
Summary
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This study reveals that CREB transcription factor binds to specific sites on the LAP/C/EBP beta gene promoter, controlling its expression during liver regeneration and in response to protein kinase A signaling.

Area of Science:

  • Molecular Biology
  • Hepatology
  • Gene Regulation

Background:

  • LAP/C/EBP beta is a transcription factor crucial for liver-specific gene expression.
  • Previous work indicated LAP/C/EBP beta mRNA modulation during liver regeneration, suggesting transcriptional control mechanisms.
  • The role of CREB (cAMP response element-binding protein) in regulating LAP/C/EBP beta transcription was investigated.

Purpose of the Study:

  • To elucidate the transcriptional mechanisms controlling LAP/C/EBP beta gene expression.
  • To identify specific DNA elements and transcription factors involved in LAP/C/EBP beta gene regulation.
  • To establish the role of CREB in LAP/C/EBP beta transcription during liver regeneration.

Main Methods:

  • Deletion analysis of the LAP/C/EBP beta 5'-flanking region using luciferase reporter constructs.

Related Experiment Videos

  • Gel shift, cross-linking, super shift, and competition assays to identify transcription factor binding.
  • Site-directed mutagenesis of identified CREB binding sites in promoter constructs.
  • Northern blot and runoff transcription assays to assess gene expression and transcription rates.
  • Western blot analysis to evaluate CREB phosphorylation status.

    • Main Results:

    • A critical region near the TATA box in the LAP/C/EBP beta promoter was identified, essential for high-level transcription.
    • Two specific binding sites for CREB were characterized within this critical promoter region.
    • Mutation of these CREB sites abolished basal promoter activity and CREB-mediated inducibility.
    • The protein kinase A pathway stimulated both reporter gene and endogenous LAP/C/EBP beta gene transcription.
    • CREB phosphorylation and LAP/C/EBP beta mRNA transcription were functionally linked during liver regeneration.

    Conclusions:

    • CREB directly binds to two sites in the LAP/C/EBP beta promoter, regulating its transcription.
    • These CREB binding sites are essential for basal and induced transcription of the LAP/C/EBP beta gene.
    • Transcriptional regulation of LAP/C/EBP beta via CREB is a key mechanism controlling gene expression under various physiological conditions, including liver regeneration.