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Related Experiment Videos

DNA binding specificity and function of retinoid X receptor alpha

J S Subauste1, R W Katz, R J Koenig

  • 1Division of Endocrinology and Metabolism, University of Michigan Medical Center, Ann Arbor 48109-0678.

The Journal of Biological Chemistry
|December 2, 1994
PubMed
Summary

Researchers identified the highest affinity binding site for Retinoid X receptors (RXRs) as GGGGTCA, surpassing the previously known AGGTCA sequence. This discovery enhances understanding of RXR transcriptional regulation.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Retinoid X receptors (RXRs) are ligand-inducible transcription factors belonging to the erbA superfamily.
  • RXRs bind to the hexameric DNA sequence AGGTCA and activate transcription upon binding to direct repeats with a one-base pair spacer in the presence of 9-cis-retinoic acid.

Purpose of the Study:

  • To determine if AGGTCA is the highest affinity binding site for RXRs.
  • To identify the optimal DNA sequence for RXR binding and transcriptional activation.

Main Methods:

  • Electrophoretic mobility shift assay combined with polymerase chain reaction to isolate high-affinity binding sequences from random DNA pools.
  • Mutational analysis and DNA footprinting to confirm the identified sequence.

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Main Results:

  • The seven-base pair sequence GGGGTCA was identified as the highest affinity RXR binding site.
  • Direct repeats of GGGGTCA function as a more potent response element than direct repeats of AGGTCA.

Conclusions:

  • The GGGGTCA sequence represents the optimal DNA binding motif for Retinoid X receptors.
  • This finding refines our understanding of RXR-mediated transcriptional regulation and response element design.