title: Detection of the argonaute protein Ago2 and microRNAs in the RNA induced silencing complex (RISC) using a monoclonal antibody.
abstract: MicroRNAs (miRNAs) are short RNA molecules responsible for post-transcriptional gene silencing by the degradation or translational inhibition of their target messenger RNAs (mRNAs). This process of gene silencing, known as RNA interference (RNAi), is mediated by highly conserved Argonaute (Ago) proteins which are the key components of the RNA induced silencing complex (RISC). In humans, Ago2 is responsible for the endonuclease cleavage of targeted mRNA and it interacts with the mRNA-binding protein GW182, which is a marker for cytoplasmic foci referred to as GW bodies (GWBs). We demonstrated that the anti-Ago2 monoclonal antibody 4F9 recognized GWBs in a cell cycle dependent manner and was capable of capturing miRNAs associated with Ago2. Since Ago2 protein is the effector protein of RNAi, anti-Ago2 monoclonal antibody may be useful in capturing functional miRNAs.
authors.affiliation: Department of Oral Biology, University of Florida, 1600 SW Archer Rd., Gainesville, FL 32610-0424, USA.
authors.forename: Keigo
authors.initials: K
authors.lastname: Ikeda
authors.forename: Minoru
authors.initials: M
authors.lastname: Satoh
authors.forename: Kaleb M
authors.initials: KM
authors.lastname: Pauley
authors.forename: Marvin J
authors.initials: MJ
authors.lastname: Fritzler
authors.forename: Westley H
authors.initials: WH
authors.lastname: Reeves
authors.forename: Edward K L
authors.initials: EK
authors.lastname: Chan
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D060565:Argonaute Proteins; D001324:Autoantigens / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D015852:Eukaryotic Initiation Factor-2; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D051379:Mice; D035683:MicroRNAs; D010448:Peptide Initiation Factors / Q000276:immunology*; D016601:RNA-Binding Proteins; D034743:RNA-Induced Silencing Complex; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: C466422:AGO2 protein, human; D000911:Antibodies, Monoclonal; D060565:Argonaute Proteins; D001324:Autoantigens; D015852:Eukaryotic Initiation Factor-2; D035683:MicroRNAs; D010448:Peptide Initiation Factors; D016601:RNA-Binding Proteins; D034743:RNA-Induced Silencing Complex; C465663:TNRC6A protein, human