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Related Experiment Videos

A consensus DNA-binding site for the androgen receptor.

P J Roche1, S A Hoare, M G Parker

  • 1Molecular Endocrinology Laboratory, Imperial Cancer Research Fund, London, United Kingdom.

Molecular Endocrinology (Baltimore, Md.)
|December 1, 1992
PubMed
Summary

Researchers identified the consensus DNA sequence for the androgen receptor (AR), finding that flanking sequences significantly impact AR binding affinity and functional activity. This discovery aids in understanding AR

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • The androgen receptor (AR) is a crucial nuclear receptor involved in various physiological processes.
  • Understanding the specific DNA sequences that AR binds to is essential for deciphering gene regulation.
  • Previous studies have identified response elements for related nuclear receptors, providing a basis for AR investigation.

Purpose of the Study:

  • To determine the consensus DNA-binding sequence for the androgen receptor (AR).
  • To investigate the influence of flanking sequences on AR binding affinity and functional activity.

Main Methods:

  • DNA-binding site selection assay using a purified AR DNA-binding domain fusion protein.
  • Gel mobility shift assays to isolate AR-oligonucleotide complexes.

Related Experiment Videos

  • Nucleotide sequencing to characterize selected DNA sites.
  • Transient transfection assays to assess functional activity of modified response elements.

    • Main Results:

    • A consensus AR-binding element was determined: 5'-GGA/TACANNNTGTTCT-3', featuring two asymmetrical 6-bp elements separated by a 3-bp spacer.
    • A slight preference for specific nucleotides in the spacer and 3'-flanking regions was observed.
    • Modifying these preferred spacer and flanking nucleotides reduced AR binding affinity by over 3-fold.
    • Changes in flanking sequences correlated with altered functional activity in reporter gene assays.

    Conclusions:

    • The consensus DNA sequence for AR binding has been elucidated.
    • Sequences flanking the core AR-binding element significantly modulate both receptor binding affinity and transcriptional activity.
    • These findings provide a deeper understanding of androgen receptor-mediated gene regulation.