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Related Experiment Videos

HuR regulates p21 mRNA stabilization by UV light.

W Wang1, H Furneaux, H Cheng

  • 1Laboratory of Biological Chemistry, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA.

Molecular and Cellular Biology
|January 11, 2000
PubMed
Summary

The RNA-binding protein HuR enhances p21 mRNA stability following UV stress. This mechanism regulates stress-induced p21 expression by increasing cytoplasmic HuR levels.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Cyclin-dependent kinase inhibitor p21 expression is induced by cellular stress.
  • Short-wavelength UV light (UVC) increases p21 mRNA stability.

Purpose of the Study:

  • To investigate the mechanisms of p21 mRNA stabilization induced by stress.
  • To identify proteins that complex with p21 mRNA upon UVC treatment.

Main Methods:

  • Utilized cytoplasmic lysates from human RKO colorectal carcinoma cells.
  • Identified RNA-binding proteins complexing with p21 mRNA using supershift assays and Western blots.
  • Assessed the role of HuR by reducing its levels using antisense HuR expression.
  • Employed luciferase reporter constructs containing the p21 3' untranslated region.

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Main Results:

  • Proteins forming complexes with p21 mRNA were inducible by UVC and other stress agents.
  • The Elav-type RNA-binding protein HuR was identified within these complexes.
  • Reduced HuR levels decreased p21 mRNA-protein complexes and reduced UVC inducibility and half-life of p21 mRNA.
  • Reduced HuR prevented UVC-mediated induction of luciferase activity in reporter constructs.

Conclusions:

  • HuR plays a critical role in regulating stress-induced p21 expression.
  • HuR enhances p21 mRNA stability, contributing to increased p21 expression under stress.
  • HuR's cytoplasmic localization is linked to its role in stabilizing p21 mRNA.