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Related Experiment Videos

Multiple cis-elements mediate shear stress-induced gene expression

J Y Shyy1, Y S Li, M C Lin

  • 1Department of Bioengineering, University of California, La Jolla 92093-0412, USA. Schien@bioeng.ucsd.edu.

Journal of Biomechanics
|December 1, 1995
PubMed
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Fluid shear stress activates immediate early (IE) genes in endothelial cells via specific DNA elements. Glucocorticoid and retinoic acid receptors can modulate this response, indicating complex transcriptional regulation.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Fluid shear stress is a critical mechanical stimulus for vascular endothelial cells.
  • Shear stress influences the expression of immediate early (IE) genes, which play roles in cellular responses.
  • Transcriptional regulation of IE genes involves specific cis-acting elements in their promoter regions.

Purpose of the Study:

  • To investigate the role of the TPA responsive element (TRE) in shear-induced monocyte chemotactic protein-1 (MCP-1) gene expression.
  • To determine if glucocorticoid and retinoic acid receptors interfere with shear stress-activated gene expression pathways.
  • To explore the involvement of multiple cis-elements in shear stress-induced IE gene expression.

Main Methods:

  • Constructing reporter gene plasmids with wild-type and mutated MCP-1 promoters linked to luciferase.

Related Experiment Videos

  • Transfecting bovine aortic endothelial cells (BAEC) with these constructs.
  • Treating cells with dexamethasone or retinoic acid prior to applying shear stress.
  • Measuring reporter gene activity to assess promoter function under different conditions.
  • Analyzing the activity of a human immunodeficiency virus long terminal repeat (HIV-LTR) construct containing a kappa B enhancer element.

    • Main Results:

    • A divergent TRE (TGACTCC) at nt -54 to -60 was identified as necessary for shear-inducibility of the MCP-1 promoter in BAEC.
    • Pretreatment with dexamethasone or retinoic acid significantly attenuated shear stress-induced MCP-1 promoter activity.
    • Glucocorticoid receptor and retinoic acid receptor appear to interfere with the shear stress-activated AP-1/TRE pathway.
    • Shear stress also activated reporter activity of the HIV-LTR construct, indicating involvement of the kappa B enhancer element.

    Conclusions:

    • The TGACTCC element is crucial for shear-induced MCP-1 gene expression in endothelial cells.
    • Glucocorticoid and retinoic acid signaling pathways can negatively regulate shear stress-induced IE gene expression.
    • Shear stress-induced immediate early gene expression is mediated through multiple cis-acting elements, including TRE and kappa B enhancer elements.