title: Peptide-Based Inhibition of miRNA-Guided Gene Silencing.
abstract: MicroRNAs (miRNAs) are a large class of small noncoding RNAs that regulate the expression of distinct target mRNAs. miRNAs are incorporated into Argonaute (AGO) proteins and guide them to their target mRNAs. Subsequently, AGO proteins recruit a member of the glycine-tryptophan-rich (GW) protein family by direct protein-protein interaction. GW proteins coordinate all downstream processes leading to robust and efficient gene silencing. A short peptide of GW proteins comprising the AGO interaction motif can be used to biochemically isolate endogenous AGO protein complexes. Furthermore, within a cell such a peptide competes with endogenous GW proteins for AGO binding and thus can be used as potent inhibitor of the miRNA pathway. Here, we describe a method that utilizes a GW-based polypeptide (T6B-assay) to validate miRNA-mRNA interactions in tissue culture systems.
authors.affiliation: Biochemistry Center Regensburg (BZR), Laboratory for RNA Biology, University of Regensburg, Universitätsstraße 31, 93053, Regensburg, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Danner
authors.affiliation: Biochemistry Center Regensburg (BZR), Laboratory for RNA Biology, University of Regensburg, Universitätsstraße 31, 93053, Regensburg, Germany.
authors.forename: Balagopal
authors.initials: B
authors.lastname: Pai
authors.affiliation: Biochemistry Center Regensburg (BZR), Laboratory for RNA Biology, University of Regensburg, Universitätsstraße 31, 93053, Regensburg, Germany.
authors.forename: Ludwig
authors.initials: L
authors.lastname: Wankerl
authors.affiliation: Biochemistry Center Regensburg (BZR), Laboratory for RNA Biology, University of Regensburg, Universitätsstraße 31, 93053, Regensburg, Germany. gunter.meister@vkl.uni-regensburg.de.
authors.forename: Gunter
authors.initials: G
authors.lastname: Meister
mesh_terms: D060565:Argonaute Proteins / Q000378:metabolism*; D020868:Gene Silencing / Q000187:drug effects; D047468:Immunoprecipitation / Q000379:methods*; D035683:MicroRNAs / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000378:metabolism / Q000627:therapeutic use*; D011485:Protein Binding; D060066:Protein Interaction Maps / Q000235:genetics; D012333:RNA, Messenger / Q000037:antagonists & inhibitors
chemical_list: D060565:Argonaute Proteins; D035683:MicroRNAs; D010455:Peptides; D012333:RNA, Messenger
keywords: Argonaute; GW proteins; TNRC6 proteins; miRNA