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Related Experiment Videos

Age-dependent decrease in the heat-inducible DNA sequence-specific binding activity in human diploid fibroblasts.

H S Choi1, Z Lin, B S Li

  • 1Department of Biological Sciences, Rutgers-State University of New Jersey, Piscataway 08855-1059.

The Journal of Biological Chemistry
|October 15, 1990
PubMed
Summary
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Aging diploid cells show reduced heat shock response due to decreased heat shock element-binding activity. This age-associated decline in transcription factor DNA-binding activity impacts heat shock gene expression.

Area of Science:

  • Molecular Biology
  • Cellular Aging
  • Gene Regulation

Background:

  • The heat shock response is crucial for cellular protection against stress.
  • Aging is associated with an attenuated heat shock response in diploid fibroblasts.
  • The molecular mechanisms underlying this age-dependent decline are not fully understood.

Purpose of the Study:

  • To investigate the regulation of a heat shock element (HSE) sequence-specific binding protein in aging IMR-90 diploid fibroblasts.
  • To determine if age-related changes in HSE binding activity contribute to the attenuated heat shock response.

Main Methods:

  • Gel retardation assay to measure HSE binding activity.
  • Scatchard analysis to assess binding affinity and capacity.
  • DNA binding assays for activating transcription factor.

Related Experiment Videos

  • Western blot analysis to determine protein molecular weight.

    • Main Results:

    • HSE binding activity was heat-inducible and transient, peaking at 1 hour.
    • This activity was significantly higher in young cells compared to old cells.
    • The difference was due to altered binding levels, not affinity, with old cells showing decreased binding capacity (Bmax).
    • An inhibitor of HSE binding was detected in old cell extracts.
    • The HSE-binding protein, identified as a heat shock gene transcription factor, had a molecular weight of 83,000 in both young and old cells.

    Conclusions:

    • There is an age-associated decrease in heat shock gene transcription factor DNA-binding activity in IMR-90 cells.
    • This decline in DNA-binding activity likely accounts for the attenuated heat shock gene expression observed in aging diploid cells.