Alternative function of a protein kinase homology domain in 2', 5'-oligoadenylate dependent RNase L.
RNase L is the 2',5'-oligoadenylate (2-5A)-dependent endoribonuclease that functions in interferon action and apoptosis. One of the intriguing, albeit unexplained, features of RNase L is its significant homology to protein kinases. Despite the homology, however, no protein kinase activity was detected during activation and RNA cleavage reactions with human RNase L. Similarly, the kinase plus ribonuclease domains of RNase L produced no detectable protein kinase activity in contrast to the phosphorylation obtained with homologous domains of the related kinase and endoribonuclease, yeast IRE1p. In addition, neither ATP nor pA(2'p5'A)3was hydrolyzed by RNase L. To further investigate the function of the kinase homology in RNase L, the conserved lysine at residue 392 in protein kinase-like domain II was replaced with an arginine residue. The resulting mutant, RNase LK392R, showed >100-fold decreases in 2-5A-dependent ribonuclease activity without reducing 2-5A- or RNA-binding activities. The greatly reduced activity of RNase LK392Rwas correlated to a defect in the ability of RNase L to dimerize. These results demonstrate a critical role for lysine 392 in the activation and dimerization of RNase L, thus suggesting that these two activities are intimately linked.
Dong
B
B
Department of Cancer Biology, NN10, The Lerner Research Institute, The Cleveland Clinic Foundation,9500 Euclid Avenue, Cleveland, OH 44195, USA.
Silverman
R H
RH
D000227:Adenine Nucleotides / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D002384:Catalysis; D020134:Catalytic Domain; D019281:Dimerization; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation; D006801:Humans; D006868:Hydrolysis; D008239:Lysine / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D009843:Oligoribonucleotides / Q000378:metabolism; D011485:Protein Binding; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D000227:Adenine Nucleotides; D009843:Oligoribonucleotides; D016601:RNA-Binding Proteins; C023505:2',5'-oligoadenylate; D000255:Adenosine Triphosphate; D011494:Protein Kinases; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease; D008239:Lysine