title: Mapping of Ago2-GW182 functional interactions.
abstract: MicroRNA (miRNA)-mediated posttranscriptional regulation of gene expression has become a major focus in understanding fine-tuning controls in many biological processes. Argonaute 2 protein (Ago2), a core component of RNA-induced silencing complex, directly binds miRNA and functions in both RNAi and miRNA pathways. GW182 is a marker protein of GW bodies (GWB, also known as mammalian P-bodies) and is known to bind the Ago2 protein. This Ago2-GW182 interaction is crucial for Ago2-miRNA-mediated translational silencing as well as the recruitment of Ago2 into GWB. Translational silencing of tethered Ago2 to a 3'UTR reporter requires GW182 for function, whereas tethered GW182 exerts a stronger repression than tethered Ago2 and does not apparently require Ago2. This chapter describes in detail the methods used in mapping Ago2-GW182 interactions.
authors.affiliation: Department of Oral Biology, University of Florida, Gainesville, FL, USA.
authors.forename: Bing
authors.initials: B
authors.lastname: Yao
authors.forename: Songqing
authors.initials: S
authors.lastname: Li
authors.forename: Shang Li
authors.initials: SL
authors.lastname: Lian
authors.forename: Marvin J
authors.initials: MJ
authors.lastname: Fritzler
authors.forename: Edward K L
authors.initials: EK
authors.lastname: Chan
mesh_terms: D060565:Argonaute Proteins; D001324:Autoantigens / Q000378:metabolism*; D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism*; D023061:Gene Order; D020868:Gene Silencing; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D021381:Protein Transport / Q000502:physiology; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism*
chemical_list: C466422:AGO2 protein, human; D060565:Argonaute Proteins; D001324:Autoantigens; D015852:Eukaryotic Initiation Factor-2; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; C465663:TNRC6A protein, human