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PMID: 14517288 Published · ppublish English Journal Article

Stabilization of urokinase and urokinase receptor mRNAs by HuR is linked to its cytoplasmic accumulation induced by activated mitogen-activated protein kinase-activated protein kinase 2.

Molecular and cellular biology ·Vol. 23 ·No. 20 ·2003-10-00 ·Pages 7177-88

Tran H, Maurer F, Nagamine Y

Abstract

The mRNAs of urokinase plasminogen activator (uPA) and its receptor, uPAR, contain instability-determining AU-rich elements (AREs) in their 3' untranslated regions. The cellular proteins binding to these RNA sequences (ARE(uPA/uPAR)) are not known. We show here that the mRNA-stabilizing factor HuR functionally interacts with these sequences. HuR stabilized an ARE(uPA)-containing RNA substrate in vitro and stabilized in HeLa Tet-off cells both endogenous uPA and uPAR mRNAs and a beta-globin reporter mRNA containing the ARE(uPA). RNAi-mediated depletion of HuR in BT-549 and MDA-MB-231 cells significantly reduced the steady-state levels of endogenous uPA and uPAR mRNAs. Furthermore, we show that a constitutively active form of mitogen-activated protein kinase-activated protein kinase 2 (MK2), MK2-EE, has an ARE-mRNA-stabilizing effect that correlates with its ability to enhance the cytoplasmic accumulation of endogenous HuR, but not in cells cotransfected with a dominant negative version of MK2, MK2-K76R. These effects were mimicked by hydrogen peroxide treatment (oxidative stress), which resulted in the phosphorylation of endogenous MK2. In addition, hydrogen peroxide treatment enhanced the cytoplasmic binding of HuR to the ARE(uPA), which was abrogated in cells transfected with MK2-K76R. These results indicate a role for HuR and MK2 in regulating the expression of uPA and uPAR genes at the posttranscriptional level.

MeSH Terms
Antigens, Surface Blotting, Northern Blotting, Western Cell Line Cell Line, Tumor Cytoplasm/metabolism Doxycycline/pharmacology ELAV Proteins ELAV-Like Protein 1 Enzyme Activation Gene Expression Regulation, Enzymologic Genes, Dominant Genetic Vectors Glutathione Transferase/metabolism HeLa Cells Humans Hydrogen Peroxide/pharmacology Intracellular Signaling Peptides and Proteins Microscopy, Fluorescence Mitogen-Activated Protein Kinases/metabolism Models, Genetic Oxidative Stress Precipitin Tests Protein Binding Protein Kinases Protein Serine-Threonine Kinases RNA/metabolism RNA, Messenger/metabolism RNA-Binding Proteins/metabolism Receptors, Cell Surface/metabolism Receptors, Urokinase Plasminogen Activator Recombinant Proteins/metabolism Signal Transduction Time Factors Transcription, Genetic Transfection Ultraviolet Rays Urokinase-Type Plasminogen Activator/metabolism
Chemicals
Antigens, Surface ELAV Proteins ELAV-Like Protein 1 ELAVL1 protein, human Intracellular Signaling Peptides and Proteins PLAUR protein, human RNA, Messenger RNA-Binding Proteins Receptors, Cell Surface Receptors, Urokinase Plasminogen Activator Recombinant Proteins RNA Hydrogen Peroxide Glutathione Transferase Protein Kinases MAP-kinase-activated kinase 2 Protein Serine-Threonine Kinases Mitogen-Activated Protein Kinases Urokinase-Type Plasminogen Activator Doxycycline
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Tran Hoanh
Friedrich Miescher Institute for Biomedical Research, Novartis Research Foundation, CH-4058 Basel, Switzerland.
Maurer Fabienne
Nagamine Yoshikuni
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.23.20.7177-7188.2003
Published
2003-10-00
Pages
7177-88
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC230330
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.23.20.7177-7188.2003 PMC Full Text View journal details
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