Involvement of hGLD‐2 in cytoplasmic polyadenylation of human p53 mRNA
Abstract:Glahder JA, Norrild B. Involvement of hGLD‐2 in cytoplasmic polyadenylation of human p53 mRNA. APMIS 2011; 119: 769–75.
Cytoplasmic polyadenylation is a post‐transcriptional mechanism regulating mRNA stability and translation. The human p53 3′‐untranslated region (3′‐UTR) contains two regions similar to cytoplasmic polyadenylation elements (CPEs) just upstream of the poly(A) hexanucleotide. Evaluation of the p53 CPE‐like elements was performed by luciferase reporter assays, qPCR, and poly(A) assays. Herein, we report the down regulation of a luciferase reporter fused to the p53 3′‐UTR, when human CPE‐binding protein 1 (hCPEB1) is overexpressed. This inhibition is partially rescued when hCPEB1fused to hGLD‐2 [a human cytoplasmic poly(A) polymerase] is overexpressed instead. The stability of a luciferase mRNA containing the p53 3′‐UTR downstream, is decreased when hCPEB1 is overexpressed as seen by qPCR. Expression of hGLD‐2 restores the mRNA stability. This is due to elongation of the poly(A) tail as seen by a PCR‐based poly(A) test and in vitro poly(A) assay. Taken together, our results suggest that hCPEB1 and hGLD‐2 are antagonizing factors regulating p53 mRNA stability.
Document Type: Research Article
Affiliations: Department of Cellular and Molecular Medicine, Panum Institute, University of Copenhagen, Copenhagen, Denmark
Publication date: November 1, 2011