Double-stranded RNA adenosine deaminases ADAR1 and ADAR2 have overlapping specificities.
Adenosine deaminases that act on RNA (ADARs) deaminate adenosines to produce inosines within RNAs that are largely double-stranded (ds). Like most dsRNA binding proteins, the enzymes will bind to any dsRNA without apparent sequence specificity. However, once bound, ADARs deaminate certain adenosines more efficiently than others. Most of what is known about the intrinsic deamination specificity of ADARs derives from analyses of Xenopus ADAR1. In addition to ADAR1, mammalian cells have a second ADAR, named ADAR2; the deamination specificity of this enzyme has not been rigorously studied. Here we directly compare the specificity of human ADAR1 and ADAR2. We find that, like ADAR1, ADAR2 has a 5' neighbor preference (A approximately U > C = G), but, unlike ADAR1, also has a 3' neighbor preference (U = G > C = A). Simultaneous analysis of both neighbor preferences reveals that ADAR2 prefers certain trinucleotide sequences (UAU, AAG, UAG, AAU). In addition to characterizing ADAR2 preferences, we analyzed the fraction of adenosines deaminated in a given RNA at complete reaction, or the enzyme's selectivity. We find that ADAR1 and ADAR2 deaminate a given RNA with the same selectivity, and this appears to be dictated by features of the RNA substrate. Finally, we observed that Xenopus and human ADAR1 deaminate the same adenosines on all RNAs tested, emphasizing the similarity of ADAR1 in these two species. Our data add substantially to the understanding of ADAR2 specificity, and aid in efforts to predict which ADAR deaminates a given editing site adenosine in vivo.
Lehmann
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KA
Department of Biochemistry and HHMI, University of Utah, 50 North Medical Drive, Room 211, Salt Lake City, Utah 84132, USA.
Bass
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D000243:Adenosine Deaminase / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D003641:Deamination; D004722:Endoribonucleases / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D008956:Models, Chemical; D008969:Molecular Sequence Data; D017393:RNA Editing; D012330:RNA, Double-Stranded / Q000737:chemistry* / Q000378:metabolism; D016601:RNA-Binding Proteins; D013045:Species Specificity; D014981:Xenopus
D012330:RNA, Double-Stranded; D016601:RNA-Binding Proteins; D004722:Endoribonucleases; C022544:ribonuclease U2; C567955:ADARB1 protein, human; D000243:Adenosine Deaminase