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Related Experiment Videos

Tonicity-regulated gene expression.

Joan D Ferraris1, Maurice B Burg

  • 1Laboratory of Kidney and Electrolyte Metabolism, National Heart Lung Blood Institute, National Institutes of Health, Department of Health and Human Services, Bethesda, Maryland, USA.

Methods in Enzymology
|September 19, 2007
PubMed
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Hypertonicity activates the TonEBP/OREBP transcription factor, increasing gene transcription. Protocols are presented to measure TonEBP/OREBP activity, nuclear localization, and DNA binding under hypertonic conditions.

Area of Science:

  • Molecular Biology
  • Cellular Physiology
  • Transcription Regulation

Background:

  • Hypertonicity is a cellular stressor that triggers adaptive responses.
  • Transcription factors play a crucial role in mediating cellular responses to osmotic stress.
  • TonEBP/OREBP is a key transcription factor activated by hypertonicity.

Purpose of the Study:

  • To present detailed protocols for analyzing TonEBP/OREBP activity.
  • To investigate the mechanisms by which hypertonicity modulates TonEBP/OREBP.
  • To provide methods for assessing TonEBP/OREBP's DNA binding and transcriptional function.

Main Methods:

  • Measurement of TonEBP/OREBP transcriptional activity.
  • Determination of TonEBP/OREBP subcellular localization (nuclear import).

Related Experiment Videos

  • Assessment of TonEBP/OREBP binding to its cognate DNA element (ORE).
  • Analysis of the transactivation domain activity of TonEBP/OREBP.

    • Main Results:

    • Hypertonicity enhances TonEBP/OREBP transcriptional activity.
    • Nuclear localization of TonEBP/OREBP is increased under hypertonic conditions.
    • TonEBP/OREBP binds to ORE sequences in a hypertonicity-dependent manner.

    Conclusions:

    • Established protocols allow for comprehensive analysis of TonEBP/OREBP regulation.
    • TonEBP/OREBP's transcriptional activity is tightly controlled by hypertonicity through multiple mechanisms.
    • Understanding TonEBP/OREBP regulation is vital for comprehending cellular responses to osmotic stress.