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Related Experiment Videos

Cell cycle-dependent glucocorticoid receptor phosphorylation and activity

J M Hu1, J E Bodwell, A Munck

  • 1Department of Physiology, Dartmouth Medical School Lebanon, New Hampshire 03756.

Molecular Endocrinology (Baltimore, Md.)
|December 1, 1994
PubMed
Summary

Cell cycle phase affects glucocorticoid receptor (GR) activity. GR phosphorylation changes explain why cells are sensitive in G1/S but resistant in G2/M, impacting disease treatment.

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Area of Science:

  • Cell Biology
  • Molecular Endocrinology
  • Biochemistry

Background:

  • Proliferating cells exhibit cell cycle-dependent sensitivity to glucocorticoid gene activation, being sensitive in late G1/S phases and resistant in G2/M.
  • Glucocorticoid receptor (GR) phosphorylation is implicated in regulating this sensitivity, with basal and hormone-induced phosphorylation patterns varying.

Purpose of the Study:

  • To investigate the cell cycle-dependent variations in GR phosphorylation and their correlation with glucocorticoid sensitivity.
  • To elucidate the role of GR phosphorylation in mediating glucocorticoid resistance during the G2/M phase.

Main Methods:

  • Utilized WCL2 cells (Chinese hamster ovary cells overexpressing GRs) to analyze GR phosphorylation.
  • Employed High-Performance Liquid Chromatography (HPLC) phosphopeptide mapping to compare GR phosphorylation sites in S and G2/M phases.

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  • Assessed GR protein levels and hormone-binding capacity across different cell cycle phases.

    • Main Results:

    • Glucocorticoid treatment failed to hyperphosphorylate GRs in G2/M but doubled phosphorylation in S phase.
    • Basal GR phosphorylation was significantly higher (nearly threefold) in G2/M compared to S phase.
    • No qualitative differences in phosphorylated sites were observed between S and G2/M phases via HPLC, suggesting a general increase in negative charge in the N-terminal domain during G2/M.
    • Increased GR protein levels correlated with enhanced hormone-binding capacity from G1 to S phase.

    Conclusions:

    • GR phosphorylation patterns are cell cycle-dependent and likely account for the observed variations in glucocorticoid sensitivity.
    • Elevated basal GR phosphorylation in G2/M may contribute to glucocorticoid resistance in this phase.
    • These findings may offer insights into glucocorticoid resistance mechanisms in inflammatory and lymphoproliferative diseases.