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Sequence Variation in the Response Element Determines Binding by the Transcription Factor p73

Ana Ramos1, Pui-Wah Tse1, Jessie Wang1

  • 1Instituto de Química, Universidad Nacional Autónoma de México (UNAM) , Circuito Exterior, Ciudad Universitaria, Mexico City, D.F. 04510, Mexico.

Biochemistry
|November 4, 2015
PubMed

Insights

The p73 transcription factor

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Transcription factors regulate gene expression by binding to specific DNA sequences.
  • The p73 transcription factor controls key cellular processes like development and apoptosis.
  • Understanding transcription factor binding is crucial for deciphering gene regulation.

Purpose of the Study:

  • To investigate how variations in DNA response element sequences affect p73 DNA-binding domain affinity.
  • To identify critical nucleotide positions within p73 response elements that dictate binding.
  • To elucidate the hierarchical nature of transcription factor-DNA interactions.

Main Methods:

  • Sedimentation velocity experiments to assess DNA-protein complex formation.
  • Fluorescence anisotropy assays to quantify DNA binding affinity.
  • Systematic sequence variations in p73 half-site response elements.

Main Results:

  • Nucleotide sequence significantly impacts p73 DNA-binding domain affinity.
  • Cytosine at the fourth position of each quarter-site is the primary determinant of binding.
  • The fifth position and purine preference in the first three positions also influence binding.
  • A hierarchical model of binding is proposed based on nucleotide position importance.

Conclusions:

  • The sequence of DNA response elements critically influences transcription factor binding affinity.
  • Specific nucleotide positions within the response element play differential roles in binding.
  • Findings support a hierarchical model for p73 transcription factor binding to DNA.

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