title: Recognition and coupling of A-to-I edited sites are determined by the tertiary structure of the RNA.
abstract: Adenosine-to-inosine (A-to-I) editing has been shown to be an important mechanism that increases protein diversity in the brain of organisms from human to fly. The family of ADAR enzymes converts some adenosines of RNA duplexes to inosines through hydrolytic deamination. The adenosine recognition mechanism is still largely unknown. Here, to investigate it, we analyzed a set of selectively edited substrates with a cluster of edited sites. We used a large set of individual transcripts sequenced by the 454 sequencing technique. On average, we analyzed 570 single transcripts per edited region at four different developmental stages from embryogenesis to adulthood. To our knowledge, this is the first time, large-scale sequencing has been used to determine synchronous editing events. We demonstrate that edited sites are only coupled within specific distances from each other. Furthermore, our results show that the coupled sites of editing are positioned on the same side of a helix, indicating that the three-dimensional structure is key in ADAR enzyme substrate recognition. Finally, we propose that editing by the ADAR enzymes is initiated by their attraction to one principal site in the substrate.
authors.affiliation: Department of Molecular Biology and Functional Genomics, Stockholm University, S-106 91 Stockholm, Sweden.
authors.forename: Mats
authors.initials: M
authors.lastname: Ensterö
authors.forename: Chammiran
authors.initials: C
authors.lastname: Daniel
authors.forename: Helene
authors.initials: H
authors.lastname: Wahlstedt
authors.forename: François
authors.initials: F
authors.lastname: Major
authors.forename: Marie
authors.initials: M
authors.lastname: Ohman
mesh_terms: D000241:Adenosine / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D007288:Inosine / Q000378:metabolism*; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D017393:RNA Editing
chemical_list: D007288:Inosine; D012313:RNA; D000241:Adenosine