Human Dicer C-terminus functions as a 5-lipoxygenase binding domain
| Autor: | Johanne Doucet, Marija Rakonjac, Andreas Hermansson, Olof Rådmark, Bengt Samuelsson, Patrick Provost, Dominique L. Ouellet, Geneviève Pépin, Vildan Dincbas-Renqvist, Isabelle Goulet, Isabelle Plante |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Lin-4 microRNA precursor
genetic processes Immunoblotting Biophysics Plasma protein binding Biochemistry Article DEAD-box RNA Helicases Structural Biology Two-Hybrid System Techniques Genetics Humans Immunoprecipitation Ribonuclease III Molecular Biology Cells Cultured Regulation of gene expression Arachidonate 5-Lipoxygenase Microscopy Confocal biology fungi RNA food and beverages Protein Structure Tertiary RNA silencing enzymes and coenzymes (carbohydrates) Microscopy Fluorescence biology.protein Mutagenesis Site-Directed Binding domain Dicer Protein Binding |
| Zdroj: | Biochimica et biophysica acta. 1789(2) |
| ISSN: | 0006-3002 |
| Popis: | Dicer is a multidomain ribonuclease III enzyme involved in the biogenesis of microRNAs (miRNAs) in the vast majority of eukaryotes. In human, Dicer has been shown to interact with cellular proteins via its N-terminal domain. Here, we demonstrate the ability of Dicer C-terminus to interact with 5-lipoxygenase (5LO), an enzyme involved in the biosynthesis of inflammatory mediators, in vitro and in cultured human cells. Yeast two-hybrid and GST binding assays delineated the smallest 5-lipoxygenase binding domain (5LObd) of Dicer to its C-terminal 140 amino acids comprising the double-stranded RNA (dsRNA) binding domain (dsRBD). The Dicer 5LObd-5LO association was disrupted upon Ala substitution of Trp residues 13, 75 and 102 in 5LO, suggesting that the Dicer 5LObd may recognize 5LO via its N-terminal C2-like domain. Whereas a catalytically active 5LObd-containing Dicer fragment was found to enhance 5LO enzymatic activity in vitro, human 5LO modified the miRNA precursor processing activity of Dicer. Providing a link between miRNA-mediated regulation of gene expression and inflammation, our results suggest that the formation of miRNAs may be regulated by 5LO in leukocytes and cancer cells expressing this lipoxygenase. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|