title: Visualization of single mRNAs reveals temporal association of proteins with microRNA-regulated mRNA.
abstract: Although many proteins are known to function in microRNA (miRNA)-based translational repression, we lack a comprehensive understanding of temporal relationships between the mRNA, miRNA and their constituent proteins. To understand the dynamics of miRNA and protein interactions, we created a synthetic inducible miRNA system in mammalian cells. By visualizing single mRNAs and observing their co-localization with proteins over time, we produced a temporal association map of miRNA-associated factors. Argonaute2, Dcp1a, hedls and Rck co-localize with miRNA-regulated mRNA after 24 h of miRNA induction, and RNAi knockdown of any one of these proteins affected the co-localization of any of the other proteins with miRNA-regulated mRNA, demonstrating that these proteins could interact with each other in a complex. We identified Argonaute2 and hedls as proteins that co-localize and interact with miRNA-regulated mRNA, indicating that processing body components are involved in long-term storage of miRNA-regulated mRNA.
authors.affiliation: Department of Systems Biology, Harvard Medical School, MA, USA.
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Shih
authors.forename: Zeev
authors.initials: Z
authors.lastname: Waks
authors.forename: Nancy
authors.initials: N
authors.lastname: Kedersha
authors.forename: Pamela A
authors.initials: PA
authors.lastname: Silver
mesh_terms: D002460:Cell Line; D017930:Genes, Reporter; D006801:Humans; D035683:MicroRNAs / Q000032:analysis / Q000378:metabolism*; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000032:analysis / Q000378:metabolism*; D034743:RNA-Induced Silencing Complex / Q000032:analysis / Q000378:metabolism
chemical_list: D035683:MicroRNAs; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D034743:RNA-Induced Silencing Complex