Chemical inhibitors destabilize HuR binding to the AU-rich element of TNF-alpha mRNA.
10.3858/emm.2009.41.11.088
- Author:
Min Ju CHAE
1
;
Hye Youn SUNG
;
Eun Hye KIM
;
Mira LEE
;
Hojoong KWAK
;
Chong Hak CHAE
;
Sunwoo KIM
;
Woong Yang PARK
Author Information
1. Department of Biomedical Sciences, Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul 110-799, Korea. wypark@snu.ac.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
anti-inflammatory agents;
ELAV-like protein 1;
lipopolysaccharides;
macrophages;
quercetin;
tumor necrosis factor-alpha
- MeSH:
*3' Untranslated Regions;
Animals;
Anti-Inflammatory Agents/*pharmacology;
Antigens, Surface/metabolism;
Antioxidants/pharmacology;
Cell Line;
Dose-Response Relationship, Drug;
Drug Evaluation, Preclinical;
Mice;
Protein Binding/drug effects;
Quercetin/*pharmacology;
RNA Stability/*drug effects;
RNA-Binding Proteins/*antagonists & inhibitors/metabolism;
Tumor Necrosis Factor-alpha/*biosynthesis
- From:Experimental & Molecular Medicine
2009;41(11):824-831
- CountryRepublic of Korea
- Language:English
-
Abstract:
Hu protein R (HuR) binds to the AU-rich element (ARE) in the 3'UTR to stabilize TNF-alpha mRNA. Here, we identified chemical inhibitors of the interaction between HuR and the ARE of TNF-alpha mRNA using RNA electrophoretic mobility gel shift assay (EMSA) and filter binding assay. Of 179 chemicals screened, we identified three with a half-maximal inhibitory concentration (IC(50)) below 10 micrometer. The IC(50) of quercetin, b-40, and b-41 were 1.4, 0.38, and 6.21 micrometer, respectively, for binding of HuR protein to TNF-alpha mRNA. Quercetin and b-40 did not inhibit binding of tristetraprolin to the ARE of TNF-alpha mRNA. When LPS-treated RAW264.7 cells were treated with quercetin and b-40, we observed decreased stability of TNF-alpha mRNA and decreased levels of secreted TNF-alpha. From these results, we could find inhibitors for the TNF-alpha mRNA stability, which might be used advantageously for both the study for post-transcriptional regulation and the discovery of new anti-inflammation drugs.
