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Related Experiment Videos

Age-related changes in DNA polymerase alpha expression.

D Busbee1, V Sylvia, G Curtin

  • 1Department of Anatomy, College of Veterinary Medicine, Texas A&M University, College Station 77843.

Experimental Gerontology
|January 1, 1989
PubMed
Summary
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As human cells age, DNA polymerase alpha activity declines, impacting DNA repair and synthesis. This decline can be reversed by specific inositol phosphates, suggesting a potential therapeutic target.

Area of Science:

  • Molecular Biology
  • Cellular Aging
  • Biochemistry

Background:

  • DNA polymerase alpha isozymes exhibit varying activity and DNA binding affinities.
  • Fetal fibroblasts possess high-activity DNA polymerase alpha (A2), while adult fibroblasts have both A2 and a low-activity form (A1).
  • The proportion of A1 increases significantly with age in human fibroblasts.

Purpose of the Study:

  • To investigate age-related changes in DNA polymerase alpha isozymes.
  • To explore the modulation of DNA polymerase alpha activity by phosphatidylinositol derivatives.
  • To assess the role of DNA polymerase alpha in age-related decline of DNA synthesis and repair.

Main Methods:

  • Purification of DNA polymerase alpha isozymes from human fibroblasts of different ages.

Related Experiment Videos

  • Enzymatic assays to determine activity and DNA binding affinity.
  • Treatment of enzymes with phosphatidylinositol-4-monophosphate (PIP) and inositol-1,4-bisphosphate (I(1,4)P2).
  • Investigation of DNA excision repair using 7,8-dihydrodiol-9,10-epoxybenzo(a)-pyrene (BPDE) in permeabilized fibroblasts.

    • Main Results:

    • The ratio of high-activity (A2) to low-activity (A1) DNA polymerase alpha decreases with age.
    • Activity and DNA binding affinity of A1 increase upon treatment with PIP or I(1,4)P2, with activity decreasing over time.
    • DNA synthesis, both scheduled and BPDE-initiated repair, declines with increasing cell age.
    • DNA polymerase alpha activity is enhanced by interaction with components of the PI phosphorylation cascade.

    Conclusions:

    • Age-related decline in DNA polymerase alpha activity and levels correlates with reduced DNA synthesis and repair capacity.
    • Activation of low-activity DNA polymerase alpha by inositol phosphates may play a role in DNA repair initiation in adult cells.
    • Phosphatidylinositol signaling pathway components can modulate DNA polymerase alpha activity, suggesting a mechanism for age-related functional changes.