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Related Experiment Videos

Negative regulation of serum-responsive enhancer elements.

M Subramaniam1, L J Schmidt, C E Crutchfield

  • 1Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905.

Nature
|July 6, 1989
PubMed
Summary
This summary is machine-generated.

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Serum growth factors induce c-fos and actin gene transcription. Protein synthesis inhibitors like cycloheximide superinduce this response, mediated by the serum response element (SRE) and its CArG box.

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Cellular Signaling

Background:

  • Serum growth factors rapidly induce transcription of proto-oncogenes like c-fos and cytoskeletal actin.
  • Protein synthesis inhibitors, such as cycloheximide, enhance this transcriptional response, a phenomenon known as superinduction.
  • The serum response element (SRE) mediates serum-induced transcription, but the mechanism of cycloheximide's superinduction effect remained unclear.

Purpose of the Study:

  • To identify the sequence elements responsible for cycloheximide-mediated superinduction of gene transcription.
  • To investigate the role of the serum response element (SRE) in mediating both serum-induced and cycloheximide-induced transcriptional changes.
  • To elucidate the regulatory mechanisms involving labile proteins and the CArG box in enhancer activity.

Related Experiment Videos

Main Methods:

  • Utilized a synthetic copy of the c-fos SRE linked to a heterologous promoter.
  • Assessed the inducibility of the promoter in the presence of cycloheximide, with and without serum.
  • Introduced mutations into the SRE to analyze their impact on both serum and cycloheximide inducibility.

Main Results:

  • A synthetic c-fos SRE conferred cycloheximide-dependent inducibility on a heterologous promoter, independent of serum.
  • Mutations within the SRE that reduced serum inducibility also impaired cycloheximide inducibility.
  • These findings suggest that labile negative regulators are involved in controlling serum-responsive enhancers.

Conclusions:

  • Serum-responsive enhancer elements are subject to negative regulation by short-lived proteins.
  • Both positive (serum) and negative (labile proteins) regulators of enhancer activity depend on a functional CArG box.
  • The CArG box plays a crucial role in mediating both serum-induced transcription and cycloheximide-dependent superinduction.