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Related Experiment Videos

Methylation-sensitive sequence-specific DNA binding by the c-Myc basic region.

G C Prendergast1, E B Ziff

  • 1Howard Hughes Medical Institute, Department of Biochemistry, New York, NY.

Science (New York, N.Y.)
|January 11, 1991
PubMed
Summary
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Researchers explored the function of the c-Myc oncoprotein in cell growth. They found that the c-Myc basic region binds DNA, influencing cell growth and cooperating with H-ras in cell transformation.

Area of Science:

  • Molecular Biology
  • Oncology
  • Cell Biology

Background:

  • The precise function of the c-Myc oncoprotein in regulating cell growth remains largely undetermined.
  • c-Myc possesses a basic region structurally similar to DNA-binding motifs found in helix-loop-helix (HLH) and leucine zipper proteins.

Purpose of the Study:

  • To investigate the DNA-binding capabilities of the c-Myc basic region.
  • To determine if the DNA-binding specificity of c-Myc is linked to its role in cell growth control and oncogenesis.

Main Methods:

  • A chimeric protein (E6) was engineered by fusing the c-Myc basic region to the HLH dimerization domain of the E12 enhancer binding factor.
  • Electrophoretic mobility shift assays (EMSAs) were employed to assess the DNA-binding activity of E6 to an E box element.

Related Experiment Videos

  • Methylation interference assays were used to probe the sequence specificity of E6 binding.
  • A rat embryo fibroblast transformation assay was conducted to evaluate the functional consequences of E6 expression in cooperation with H-ras.

    • Main Results:

    • The chimeric E6 protein specifically bound to an E box DNA element in a manner dependent on the c-Myc basic region's integrity.
    • Methylation of a CpG site within the E box specifically inhibited E6 binding, indicating sequence-specific recognition.
    • Expression of functional E6, but not a DNA-binding mutant, suppressed the cooperative transformation of rat embryo fibroblasts by c-myc and H-ras.

    Conclusions:

    • The c-Myc basic region possesses intrinsic DNA-binding activity with specific recognition properties.
    • The DNA-binding function of c-Myc is implicated in its ability to cooperate with other oncogenes like H-ras in cellular transformation.
    • These findings provide insights into the molecular mechanisms underlying c-Myc's role in cell growth and cancer.