Altered RNA editing in mice lacking ADAR2 autoregulation.
ADAR2 is a double-stranded-RNA-specific adenosine deaminase involved in the editing of mammalian RNAs by the site-selective conversion of adenosine to inosine. Previous studies from our laboratory have demonstrated that ADAR2 can modify its own pre-mRNA to create a proximal 3' splice site containing a noncanonical adenosine-inosine dinucleotide. Alternative splicing to this proximal acceptor adds 47 nucleotides to the mature ADAR2 transcript, thereby resulting in the loss of functional ADAR2 protein expression due to premature translation termination in an alternate reading frame. To examine whether the editing of ADAR2 transcripts represents a negative autoregulatory strategy to modulate ADAR2 protein expression, we have generated genetically modified mice in which the ability of ADAR2 to edit its own pre-mRNA has been selectively ablated by deletion of a critical sequence (editing site complementary sequence [ECS]) required for adenosine-to-inosine conversion. Here we demonstrate that ADAR2 autoediting and subsequent alternative splicing are abolished in homozygous deltaECS mice and that ADAR2 protein expression is increased in numerous tissues compared to wild-type animals. The observed increases in ADAR2 protein expression correlate with the extent of ADAR2 autoediting observed with wild-type tissues and correspond to increases in the editing of ADAR2 substrates, indicating that ADAR2 autoediting is a key regulator of ADAR2 protein expression and activity in vivo.
Feng
Yi
Y
Department of Pharmacology, Vanderbilt University School of Medicine, 465 21st Avenue South, 8160 Medical Research Building 3, Nashville, Tennessee 37232-8548, USA.
Sansam
Christopher L
CL
Singh
Minati
M
Emeson
Ronald B
RB
D000243:Adenosine Deaminase / Q000096:biosynthesis* / Q000235:genetics; D017398:Alternative Splicing / Q000502:physiology; D000818:Animals; D015870:Gene Expression; D008297:Male; D051379:Mice; D008817:Mice, Mutant Strains; D009154:Mutation; D017393:RNA Editing; D012333:RNA, Messenger / Q000096:biosynthesis; D016601:RNA-Binding Proteins
D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C567955:ADARB1 protein, human; D000243:Adenosine Deaminase