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Related Experiment Videos

Functional consensus for mammalian osmotic response elements.

J D Ferraris1, C K Williams, A Ohtaka

  • 1Osmotic Regulation Section, Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892-1603, USA. jdf@helix.nih.gov

The American Journal of Physiology
|March 10, 1999
PubMed
Summary

Mammalian cells adapt to high salt by accumulating sorbitol via aldose reductase (AR) gene transcription. Researchers defined the key DNA sequence (osmotic response element) controlling this adaptation, aiding discovery of other osmotically regulated genes.

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Area of Science:

  • Molecular Biology
  • Cellular Physiology
  • Genetics

Background:

  • Organisms universally adapt to hyperosmotic stress by accumulating organic osmolytes.
  • Mammalian renal medulla cells face high salt concentrations, accumulating sorbitol via aldose reductase (AR) gene induction.
  • Previous work identified a minimal osmotic response element (ORE) in the rabbit AR gene.

Purpose of the Study:

  • To functionally characterize the mammalian osmotic response element (ORE) controlling aldose reductase gene transcription.
  • To determine the precise DNA sequence required for hyperosmotic response.
  • To establish a consensus sequence for mammalian OREs.

Main Methods:

  • Systematic base-pair substitution within the ORE sequence.
  • Reporter gene assays to measure transcriptional activity.

Related Experiment Videos

  • Electrophoretic mobility shift assays (EMSA) to assess DNA-binding activity.

    • Main Results:

    • Functional analysis of all possible single-base substitutions within the ORE was performed.
    • EMSA confirmed binding activity correlated with osmotic response.
    • A functional consensus sequence for the mammalian ORE was determined: NGGAAAWDHMC(N).

    Conclusions:

    • The study precisely defines the functional consensus sequence of the mammalian ORE.
    • This detailed understanding of the ORE will facilitate the identification of novel osmotically regulated genes.
    • The findings advance our knowledge of cellular adaptation mechanisms to osmotic stress.