title: MicroRNA miR-324-3p induces promoter-mediated expression of RelA gene.
abstract: MicroRNAs (miRNAs) are known to repress translation by binding to the 3'UTRs of mRNAs. Using bioinformatics, we recently reported that several miRNAs also have target sites in DNA particularly in the promoters of the protein-coding genes. To understand the functional significance of this phenomenon, we tested the effects of miR-324-3p binding to RelA promoter. In PC12 cells, co-transfection with premiR-324-3p induced a RelA promoter plasmid in a dose-dependent manner and this effect was lost when the miR-324-3p binding site in the promoter was mutated. PremiR-324-3p transfection also significantly induced the endogenous RelA mRNA and protein expression in PC12 cells. Furthermore, transfection with premiR-324-3p increased the levels of cleaved caspase-3 which is a marker of apoptosis. Importantly, the miR-324-3p effects were Ago2 mediated as Ago2 knockdown prevented RelA expression and cleavage of caspase-3. Thus, our studies show that miRNA-mediated transcriptional activation can be seen in PC12 cells which are neural in origin.
authors.affiliation: Department of Neurological Surgery, University of Wisconsin, Madison, Wisconsin, United States of America ; Neuroscience Training Program, University of Wisconsin, Madison, Wisconsin, United States of America.
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Dharap
authors.forename: Courtney
authors.initials: C
authors.lastname: Pokrzywa
authors.forename: Shruthi
authors.initials: S
authors.lastname: Murali
authors.forename: Gopal
authors.initials: G
authors.lastname: Pandi
authors.forename: Raghu
authors.initials: R
authors.lastname: Vemuganti
mesh_terms: D000818:Animals; D060565:Argonaute Proteins; D001483:Base Sequence; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics*; D016716:PC12 Cells; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D051996:Transcription Factor RelA / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation / Q000235:genetics*
chemical_list: C582332:Ago2 protein, rat; D060565:Argonaute Proteins; D015852:Eukaryotic Initiation Factor-2; C576416:MIRN324 microRNA, rat; D008565:Membrane Proteins; D035683:MicroRNAs; D012333:RNA, Messenger; C498145:Rela protein, rat; D051996:Transcription Factor RelA