Chemical inhibitors destabilize HuR binding to the AU-rich element of TNF-alpha mRNA.
Hu protein R (HuR) binds to the AU-rich element (ARE) in the 3UTR 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 microM. The IC(50) of quercetin, b-40, and b-41 were 1.4, 0.38, and 6.21 microM, 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.
Chae
Min-Ju
MJ
Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea.
Sung
Hye Youn
HY
Kim
Eun-Hye
EH
Lee
Mira
M
Kwak
Hojoong
H
Chae
Chong Hak
CH
Kim
Sunwoo
S
Park
Woong-Yang
WY
D020413:3' Untranslated Regions; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D000954:Antigens, Surface / Q000378:metabolism; D000975:Antioxidants / Q000494:pharmacology; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D051959:ELAV Proteins; D000067780:ELAV-Like Protein 1; D051379:Mice; D011485:Protein Binding / Q000187:drug effects; D011794:Quercetin / Q000494:pharmacology*; D020871:RNA Stability / Q000187:drug effects*; D016601:RNA-Binding Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D020413:3' Untranslated Regions; D000893:Anti-Inflammatory Agents; D000954:Antigens, Surface; D000975:Antioxidants; D051959:ELAV Proteins; D000067780:ELAV-Like Protein 1; C098770:ELAVL1 protein, human; D016601:RNA-Binding Proteins; D014409:Tumor Necrosis Factor-alpha; D011794:Quercetin