title: FRET analysis of in vivo dimerization by RNA-editing enzymes.
abstract: Members of the ADAR (adenosine deaminase that acts on RNA) enzyme family catalyze the hydrolytic deamination of adenosine to inosine within double-stranded RNAs, a poorly understood process that is critical to mammalian development. We have performed fluorescence resonance energy transfer experiments in mammalian cells transfected with fluorophore-bearing ADAR1 and ADAR2 fusion proteins to investigate the relationship between these proteins. These studies conclusively demonstrate the homodimerization of ADAR1 and ADAR2 and also show that ADAR1 and ADAR2 form heterodimers in human cells. RNase treatment of cells expressing these fusion proteins changes their localization but does not affect dimerization. Taken together these results suggest that homo- and heterodimerization are important for the activity of ADAR family members in vivo and that these associations are RNA independent.
authors.affiliation: Department of Biochemistry, University of Alberta, Edmonton, Alberta T6G 2H7, Canada.
authors.forename: Kaari A
authors.initials: KA
authors.lastname: Chilibeck
authors.forename: Tao
authors.initials: T
authors.lastname: Wu
authors.forename: Chao
authors.initials: C
authors.lastname: Liang
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Schellenberg
authors.forename: Emily M
authors.initials: EM
authors.lastname: Gesner
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Lynch
authors.forename: Andrew M
authors.initials: AM
authors.lastname: MacMillan
mesh_terms: D000243:Adenosine Deaminase / Q000737:chemistry*; D019281:Dimerization; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D012313:RNA / Q000737:chemistry*; D017393:RNA Editing; D016601:RNA-Binding Proteins; D012260:Ribonucleases / Q000737:chemistry; D014162:Transfection
chemical_list: D016601:RNA-Binding Proteins; D012313:RNA; D012260:Ribonucleases; C567955:ADARB1 protein, human; D000243:Adenosine Deaminase