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PMID: 20571027 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Mechanism of regulation of bcl-2 mRNA by nucleolin and A+U-rich element-binding factor 1 (AUF1).

The Journal of biological chemistry ·Vol. 285 ·No. 35 ·2010-08-27 ·Pages 27182-27191

Ishimaru D, Zuraw L, Ramalingam S, Sengupta TK, Bandyopadhyay S, Reuben A, Fernandes DJ, Spicer EK

Abstract

The antiapoptotic Bcl-2 protein is overexpressed in a variety of cancers, particularly leukemias. In some cell types this is the result of enhanced stability of bcl-2 mRNA, which is controlled by elements in its 3'-untranslated region. Nucleolin is one of the proteins that binds to bcl-2 mRNA, thereby increasing its half-life. Here, we examined the site on the bcl-2 3'-untranslated region that is bound by nucleolin as well as the protein binding domains important for bcl-2 mRNA recognition. RNase footprinting and RNA fragment binding assays demonstrated that nucleolin binds to a 40-nucleotide region at the 5' end of the 136-nucleotide bcl-2 AU-rich element (ARE(bcl-2)). The first two RNA binding domains of nucleolin were sufficient for high affinity binding to ARE(bcl-2). In RNA decay assays, ARE(bcl-2) transcripts were protected from exosomal decay by the addition of nucleolin. AUF1 has been shown to recruit the exosome to mRNAs. When MV-4-11 cell extracts were immunodepleted of AUF1, the rate of decay of ARE(bcl-2) transcripts was reduced, indicating that nucleolin and AUF1 have opposing roles in bcl-2 mRNA turnover. When the function of nucleolin in MV-4-11 cells was impaired by treatment with the nucleolin-targeting aptamer AS1411, association of AUF1 with bcl-2 mRNA was increased. This suggests that the degradation of bcl-2 mRNA induced by AS1411 results from both interference with nucleolin protection of bcl-2 mRNA and recruitment of the exosome by AUF1. Based on our findings, we propose a model that illustrates the opposing roles of nucleolin and AUF1 in regulating bcl-2 mRNA stability.

MeSH Terms
3' Untranslated Regions Aptamers, Nucleotide HeLa Cells Heterogeneous Nuclear Ribonucleoprotein D0 Heterogeneous-Nuclear Ribonucleoprotein D/genetics,metabolism Humans Leukemia/genetics,mortality Models, Biological Oligodeoxyribonucleotides/pharmacology Phosphoproteins/antagonists & inhibitors,genetics,metabolism Protein Binding/drug effects,physiology Protein Structure, Tertiary Proto-Oncogene Proteins c-bcl-2/biosynthesis,genetics RNA Stability/drug effects,physiology RNA, Neoplasm/genetics,metabolism RNA-Binding Proteins/antagonists & inhibitors,genetics,metabolism
Chemicals
3' Untranslated Regions AGRO 100 Aptamers, Nucleotide HNRNPD protein, human Heterogeneous Nuclear Ribonucleoprotein D0 Heterogeneous-Nuclear Ribonucleoprotein D Oligodeoxyribonucleotides Phosphoproteins Proto-Oncogene Proteins c-bcl-2 RNA, Neoplasm RNA-Binding Proteins nucleolin
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Ishimaru Daniella
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina 29425.
Zuraw Lisa
Department of Chemistry, The Citadel, Charleston, South Carolina 29409.
Ramalingam Sivakumar
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina 29425.
Sengupta Tapas K
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina 29425.
Bandyopadhyay Sumita
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina 29425.
Reuben Adrian
Division of Gastroenterology and Hepatology, Department of Medicine, Medical University of South Carolina, Charleston, South Carolina 29425.
Fernandes Daniel J
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina 29425.
Spicer Eleanor K
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina 29425. Electronic address: spicer@musc.edu.
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M109.098830
Email
spicer@musc.edu
Published
2010-08-27
Epub
2010-00-22
Pages
27182-27191
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2930717
Subset
IM
Grants
NCI NIH HHS · R01 CA087553 · United States
NCI NIH HHS · R01 CA109254 · United States
NCI NIH HHS · CA 87553 · United States
NCI NIH HHS · CA109254 · United States
External Links
PubMed source DOI: 10.1074/jbc.M109.098830 PMC Full Text View journal details
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