Adenosine deaminase acting on RNA 1 accelerates cell cycle through increased translation and activity of cyclin-dependent kinase 2.
Adenosine-to-inosine RNA editing has been recently implicated in the pathogenesis of inflammation through the upregulation of the editase adenosine deaminase acting on RNA 1 (ADAR1). Because cell proliferation is a key feature of the inflammatory process, the present study tested the hypothesis that overexpression of ADAR1 accelerates cell cycle. To that end, human embryonic kidney 293 cells were transiently transfected with ADAR1 or vector, and cell cycle was evaluated by fluorescence-activated cell sorter. Overexpression of wild-type ADAR1 decreased the proportion of G0-G1 cells (-19%, P<0.01, n=3), increased the percentage of S phase cells (+19%, P<0.01, n=3), and did not change the ratio of cells residing in the G2-M phase (n=3). This finding was supported by three observations. First, there was a parallel production in ADAR1-transfected cells of cyclin-dependent kinase (Cdk) 2 and cyclin A, a pivotal protein complex upregulated at the G1-S phase checkpoint, and of [p]-Histone H1, a marker of Cdk2 activity (+102%, P<0.01, n=3). Second, ADAR1-transfected cells displayed higher activity of the proliferation marker, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide. Third, using anti-ADAR1 antibody, direct binding of ADAR1 to Cdk2 messenger RNA was demonstrated in ADAR1-transfected cells by protein-RNA cross-linking and immunoprecipitation (+974%, P<0.01, n=3). Finally, causal relationships between ADAR1 and Cdk2 were confirmed by a study with the Cdk2 inhibitor, kenpaullone, which prevented the ADAR1-induced shift from the G0-G1 to the S phase. Taken together, these data show that ADAR1 increases cell cycle by shifting cells from the G0-G1 to the S phase through the upregulation of Cdk2.
Zhang
Fangming
F
Section of Trauma, Surgical Critical Care, and Surgical Emergencies, Yale University School of Medicine, New Haven, CT 06520, USA.
Rabinovici
Reuven
R
D000243:Adenosine Deaminase / Q000378:metabolism*; D002453:Cell Cycle / Q000235:genetics; D002460:Cell Line; D051357:Cyclin-Dependent Kinase 2 / Q000096:biosynthesis*; D006801:Humans; D014176:Protein Biosynthesis / Q000235:genetics; D016601:RNA-Binding Proteins; D014162:Transfection; D015854:Up-Regulation / Q000235:genetics
D016601:RNA-Binding Proteins; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; C567955:ADARB1 protein, human; D000243:Adenosine Deaminase