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Isolation of Proteins Associated with the DNA-Bound Estrogen Receptor α

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Molecular Endocrinology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 590))

Abstract

Regulating gene expression is a complex process requiring the interaction of multiple transcription factors with their cognate recognition sequences. While these DNA-bound transcription factors are the primary drivers of gene expression, the capacity of a transcription factor to alter gene expression is tempered by its association with a host of coregulatory proteins that are recruited to the DNA-bound transcription factor. We have developed a novel approach to isolate large complexes of proteins associated with the DNA-bound estrogen receptor α (ERα) using an agarose-based electrophoretic mobility shift assay (EMSA). This method should be readily adapted to a variety of cultured cell lines, DNA sequences, and transcription factors and has the potential to provide valuable information about a wide variety of regulatory proteins involved in influencing gene expression.

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Acknowledgements

We are indebted to J. Yates and coworkers for their mass spectrometry analysis of the proteins isolated. This work was supported by NIH grant R01 DK 53884 (to AMN). JRS-N was supported by an NIH Reproductive Biology Program Training Grant (T32 HD07028).

Author information

Authors and Affiliations

  1. Department of Molecular and Integrative Physiology, University of Illinois, Urbana, IL, USA

    Jennifer R. Schultz-Norton, Yvonne S. Ziegler & Ann M. Nardulli

  2. Department of Biochemistry, University of Illinois, Urbana, IL, USA

    Varsha S. Likhite

Authors
  1. Jennifer R. Schultz-Norton
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  2. Yvonne S. Ziegler
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  3. Varsha S. Likhite
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  4. Ann M. Nardulli
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Editor information

Editors and Affiliations

  1. College of Medicine, University of Kentucky, Rose Street 800, Lexington, 40536-0298, U.S.A.

    Ok-Kyong Park-Sarge

  2. Dept. Obstetrics & Gynecology, University of Kentucky, Rose Street 800, Lexington, 40536-0293, U.S.A.

    Thomas E. Curry

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Schultz-Norton, J.R., Ziegler, Y.S., Likhite, V.S., Nardulli, A.M. (2009). Isolation of Proteins Associated with the DNA-Bound Estrogen Receptor α. In: Park-Sarge, OK., Curry, T. (eds) Molecular Endocrinology. Methods in Molecular Biology, vol 590. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-378-7_13

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  • DOI: https://doi.org/10.1007/978-1-60327-378-7_13

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60327-377-0

  • Online ISBN: 978-1-60327-378-7

  • eBook Packages: Springer Protocols

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