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Related Experiment Videos

Regulation of A-raf expression

J E Lee1, T W Beck, L Wojnowski

  • 1Biological Carcinogenesis and Development Program, Program Resources, Inc.@Dyn Corp., Frederick, Maryland, USA

Oncogene
|April 18, 1996
PubMed
Summary
This summary is machine-generated.

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The A-raf proto-oncogene

Area of Science:

  • Molecular Biology
  • Cell Signaling
  • Cancer Genetics

Background:

  • A-raf is a proto-oncogene encoding a cytoplasmic protein serine/threonine kinase.
  • It plays a role in cell growth, development, and Ras signaling pathways.
  • A-raf transcripts are primarily found in mouse urogenital tissues.

Purpose of the Study:

  • To investigate the functional role of glucocorticoid response elements (GREs) in the human A-raf promoter.
  • To determine if glucocorticoid receptors regulate A-raf gene expression.

Main Methods:

  • Transfection of HeLa cells with A-raf promoter/reporter constructs and glucocorticoid receptor expression vectors.
  • Dexamethasone induction assays to measure promoter activity.
  • Electrophoretic mobility shift assays (EMSAs) to analyze protein-DNA interactions.

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Main Results:

  • Dexamethasone treatment induced A-raf promoter activity up to fivefold in constructs containing all three GRE motifs.
  • Mutations in GREs reduced or eliminated dexamethasone-dependent induction.
  • EMSAs confirmed binding of the glucocorticoid receptor DNA binding domain to GRE-2 and GRE-3 with varying affinities.
  • A distinct protein-DNA complex with GRE-1 was observed, suggesting interaction with non-glucocorticoid receptor proteins.

Conclusions:

  • The A-raf promoter is partially regulated by glucocorticoid receptor family members.
  • Specific GREs in the A-raf promoter bind the glucocorticoid receptor.
  • A novel protein interacts with GRE-1, indicating complex regulation of A-raf expression in urogenital tissues.