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Yeast As a Chassis for Developing Functional Assays to Study Human P53
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Yeast As a Chassis for Developing Functional Assays to Study Human P53

Published on: August 4, 2019

Involvement of hGLD-2 in cytoplasmic polyadenylation of human p53 mRNA

Jacob A Glahder1, Bodil Norrild

  • 1Department of Cellular and Molecular Medicine, Panum Institute, University of Copenhagen, Denmark.

APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica
|October 15, 2011
PubMed

Insights

Human CPE-binding protein 1 (hCPEB1) and hGLD-2 oppositely regulate p53 mRNA stability. Overexpressing hCPEB1 decreases p53 mRNA stability, while hGLD-2 expression restores it by elongating the poly(A) tail.

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Post-transcriptional Modification

Background:

  • Cytoplasmic polyadenylation is a key mechanism controlling mRNA fate.
  • The human p53 3'-untranslated region (3'-UTR) possesses sequences resembling cytoplasmic polyadenylation elements (CPEs).

Purpose of the Study:

  • To investigate the role of human CPE-binding protein 1 (hCPEB1) and its interaction with hGLD-2 in regulating p53 mRNA stability.
  • To elucidate the impact of hCPEB1 and hGLD-2 on p53 mRNA polyadenylation and stability.

Main Methods:

  • Luciferase reporter assays were employed to assess p53 3'-UTR activity.
  • Quantitative PCR (qPCR) was used to measure mRNA stability.
  • Poly(A) assays, including PCR-based and in vitro methods, were performed to analyze poly(A) tail length.

Main Results:

  • Overexpression of hCPEB1 led to the downregulation of a luciferase reporter linked to the p53 3'-UTR.
  • hCPEB1 overexpression decreased the stability of p53 mRNA, evidenced by qPCR analysis.
  • Co-expression with hGLD-2 partially rescued reporter activity and restored mRNA stability, attributed to poly(A) tail elongation.

Conclusions:

  • hCPEB1 acts as an inhibitory factor on p53 mRNA stability and translation.
  • hGLD-2, a cytoplasmic poly(A) polymerase, counteracts hCPEB1's effect by elongating the poly(A) tail.
  • hCPEB1 and hGLD-2 function antagonistically to modulate p53 mRNA stability, highlighting a novel regulatory pathway.

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