title: A structural determinant required for RNA editing.
abstract: RNA editing by adenosine deaminases acting on RNAs (ADARs) can be both specific and non-specific, depending on the substrate. Specific editing of particular adenosines may depend on the overall sequence and structural context. However, the detailed mechanisms underlying these preferences are not fully understood. Here, we show that duplex structures mimicking an editing site in the Gabra3 pre-mRNA unexpectedly fail to support RNA editing at the Gabra3 I/M site, although phylogenetic analysis suggest an evolutionarily conserved duplex structure essential for efficient RNA editing. These unusual results led us to revisit the structural requirement for this editing by mutagenesis analysis. In vivo nuclear injection experiments of mutated editing substrates demonstrate that a non-conserved structure is a determinant for editing. This structure contains bulges either on the same or the strand opposing the edited adenosine. The position of these bulges and the distance to the edited base regulate editing. Moreover, elevated folding temperature can lead to a switch in RNA editing suggesting an RNA structural change. Our results indicate the importance of RNA tertiary structure in determining RNA editing.
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zhejiang University (Zijingang Campus), Hangzhou, Zhejiang, ZJ310058, PR of China.
authors.forename: Nan
authors.initials: N
authors.lastname: Tian
authors.forename: Yun
authors.initials: Y
authors.lastname: Yang
authors.forename: Nora
authors.initials: N
authors.lastname: Sachsenmaier
authors.forename: Dominik
authors.initials: D
authors.lastname: Muggenhumer
authors.forename: Jingpei
authors.initials: J
authors.lastname: Bi
authors.forename: Christina
authors.initials: C
authors.lastname: Waldsich
authors.forename: Michael F
authors.initials: MF
authors.lastname: Jantsch
authors.forename: Yongfeng
authors.initials: Y
authors.lastname: Jin
mesh_terms: D000818:Animals; D002417:Cattle; D004285:Dogs; D019143:Evolution, Molecular; D051379:Mice; D009690:Nucleic Acid Conformation; D017393:RNA Editing; D012322:RNA Precursors / Q000737:chemistry / Q000378:metabolism; D012330:RNA, Double-Stranded / Q000737:chemistry; D012333:RNA, Messenger / Q000737:chemistry* / Q000378:metabolism; D011963:Receptors, GABA-A / Q000235:genetics; D013696:Temperature
chemical_list: C546258:Gabra3 protein, mouse; D012322:RNA Precursors; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D011963:Receptors, GABA-A