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Detecting the Ligand-binding Domain Dimerization Activity of Estrogen Receptor Alpha Using the Mammalian Two-Hybrid Assay
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Age-dependent decrease in the interaction of beta-tubulin with estrogen receptor alpha transactivation domain in

Swati Ghosh1, M K Thakur

  • 1Biochemistry and Molecular Biology Laboratory, Department of Zoology, Banaras Hindu University, Varanasi 221005, India.

Neuroscience Letters
|September 1, 2009
PubMed
Summary

The 50 kDa protein interacting with estrogen receptor alpha (ERα) in the mouse brain is identified as beta-tubulin. Its interaction varies with age and sex, potentially influencing estrogen-mediated brain functions.

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Endocrinology

Background:

  • Previous studies identified four mouse brain proteins interacting with estrogen receptor alpha (ERα) transactivation domain (TAD).
  • A 68 kDa protein exhibited age- and sex-dependent changes in its interaction with ERα-TAD.

Purpose of the Study:

  • To identify the 50 kDa protein interacting with ERα-TAD.
  • To investigate age- and sex-dependent changes in the interaction between the 50 kDa protein and ERα-TAD in the mouse brain.

Main Methods:

  • Protein identification using biochemical assays.
  • Analysis of protein-protein interactions between ERα-TAD and identified proteins.
  • Assessment of protein expression levels.

Main Results:

  • The 50 kDa protein was identified as beta-tubulin, a microtubule-associated protein.
  • Beta-tubulin binds to the activation function 1 (AF-1) region of ERα-TAD.
  • Interaction between ERα-TAD and beta-tubulin was higher in adult females compared to old females and adult males.
  • Beta-tubulin expression showed no significant changes with age or sex.

Conclusions:

  • Beta-tubulin is a novel interactor of ERα-TAD involved in estrogen signaling.
  • Age- and sex-dependent alterations in beta-tubulin and ERα-TAD interaction may influence estrogen-mediated brain functions during aging.