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

TY - JOUR

T1 - Involvement of hGLD-2 in cytoplasmic polyadenylation of human p53 mRNA

AU - Glahder, Jacob-Andreas Harald

AU - Norrild, Bodil

N1 - © 2011 The Authors. APMIS © 2011 APMIS.

PY - 2011/11

Y1 - 2011/11

N2 - 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.

AB - 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.

KW - 3' Untranslated Regions

KW - Cytoplasm

KW - Humans

KW - Polyadenylation

KW - RNA Stability

KW - RNA, Messenger

KW - Transcription Factors

KW - Tumor Suppressor Protein p53

KW - mRNA Cleavage and Polyadenylation Factors

U2 - 10.1111/j.1600-0463.2011.02804.x

DO - 10.1111/j.1600-0463.2011.02804.x

M3 - Journal article

C2 - 21995630

SN - 0903-465X

VL - 119

SP - 769

EP - 775

JO - APMIS. Supplementum

JF - APMIS. Supplementum

IS - 11

ER -