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Cloning of a cDNA encoding a human protein which binds a sequence in the c-myc gene similar to the

Y Z Stasiv1, T D Mashkova, B K Chernov

  • 1Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow.

Gene
|August 5, 1994
PubMed

Insights

Researchers identified a novel human protein that binds to the c-myc promoter. This protein, named IRLB (interferon-response-like binding), specifically recognizes an element crucial for c-myc gene regulation.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • The human c-myc gene is a critical proto-oncogene involved in cell growth and proliferation.
  • Understanding the regulation of c-myc expression is essential for comprehending cancer development.
  • Specific DNA-binding proteins play a key role in controlling gene transcription.

Purpose of the Study:

  • To isolate and characterize a protein that binds to a specific element in the human c-myc promoter.
  • To identify the DNA-binding domain and specificity of the isolated protein.
  • To elucidate the role of this protein in c-myc gene regulation.

Main Methods:

  • Screening of a human fibroblast lambda gt11 phage library using a specific oligonucleotide probe.
  • Construction of an Escherichia coli strain expressing the cloned fusion protein.
  • Analysis of protein-DNA interactions using DNase footprinting and oligonucleotide-binding specificity assays.

Main Results:

  • A human cDNA clone encoding a c-myc promoter-binding protein (IRLB) was successfully isolated.
  • The deduced amino acid sequence revealed a putative DNA-binding domain.
  • DNase footprinting and binding assays confirmed that IRLB specifically recognizes the ISRE-like element in the c-myc P2 promoter region.

Conclusions:

  • A novel human protein, IRLB, that binds to the c-myc promoter has been identified.
  • IRLB specifically recognizes an interferon-stimulated response element (ISRE)-like sequence within the c-myc P2 promoter.
  • This finding contributes to understanding the molecular mechanisms regulating human c-myc gene expression.

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