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Related Experiment Videos

Artificial steroid hormone response element generated by dam-methylation.

M Truss1, J Bartsch, G Chalepakis

  • 1Institut für Molekularbiologie und Tumorforschung, Marburg, Germany.

Nucleic Acids Research
|April 11, 1992
PubMed
Summary
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Bacterial cloning can alter DNA sequences, leading to inaccurate results. Using dam- bacterial strains is crucial for correctly identifying DNA sequences, especially for hormone responsive elements.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Steroid hormone receptors bind to specific DNA sequences called palindromic response elements.
  • Bacterial DNA methylation can interfere with the accurate identification of these elements.
  • The dam+ bacterial strain methylates adenine residues, potentially causing artifacts.

Purpose of the Study:

  • To investigate the impact of adenine methylation in dam+ bacterial strains on the function of hormone responsive elements.
  • To determine the significance of hydrophobic interactions in receptor-DNA binding.
  • To highlight the importance of using appropriate bacterial strains for DNA sequence analysis.

Main Methods:

  • Studying the interaction of steroid hormone receptors with modified palindromic response elements.

Related Experiment Videos

  • Utilizing dam+ and dam- bacterial strains for DNA amplification and cloning.
  • Comparing the binding affinity of receptors to methylated and unmethylated DNA sequences.

    • Main Results:

    • Adenine methylation at the N-6 position in dam+ bacteria can create artifactual functional hormone responsive elements.
    • A specific T to A substitution in a mutant palindrome gains functional affinity only when amplified in dam+ bacteria.
    • Methylated mutant palindromes show similar receptor binding affinity as consensus elements, unlike unmethylated counterparts.

    Conclusions:

    • Hydrophobic interactions between receptors and DNA's major groove are critical for sequence discrimination.
    • Artifactual results can arise from adenine methylation during cloning in dam+ bacteria.
    • Employing dam- bacterial strains is essential for accurate identification of cis-acting DNA elements.