Regulation of phosphorylation of aminoacyl-tRNA synthetases in the high molecular weight core complex in reticulocytes.
Five aminoacyl-tRNA synthetases found in the high molecular weight core complex were phosphorylated in rabbit reticulocytes following labeling with 32P. The synthetases were isolated by affinity chromatography on tRNA-Sepharose followed by immunoprecipitation. The five synthetases phosphorylated were the glutamyl-, glutaminyl-, lysyl-, and aspartyl-tRNA synthetases and, to a lesser extent, the methionyl-tRNA synthetase. In addition, a 37,000-dalton protein, associated with the synthetase complex and tentatively identified as casein kinase I, was also phosphorylated in intact cells. Phosphoamino acid analysis of the proteins indicated all of the phosphate was on seryl residues. Incubation of reticulocytes with 32P in the presence of 8-bromo-cAMP and 3-isobutyl-1-methylxanthine resulted in a 6-fold increase in phosphorylation of the glutaminyl-tRNA synthetase and a 2-fold increase in phosphorylation of the aspartyl-tRNA synthetase. When the high molecular weight core complex was isolated by gel filtration/affinity chromatography, the profile of phosphorylation was similar to that observed by immunoprecipitation with a 9- and 3-fold stimulation of the glutaminyl- and aspartyl tRNA-synthetase, respectively. From this data it was concluded that the increased phosphorylation of the glutaminyl- and aspartyl-tRNA synthetases obtained with 8-bromo-cAMP did not appear to be involved in dissociation of the high molecular weight core complex.
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000604:Amino Acyl-tRNA Synthetases / Q000097:blood*; D000818:Animals; D001226:Aspartate-tRNA Ligase / Q000378:metabolism; D005975:Glutamate-tRNA Ligase / Q000097:blood; D008250:Lysine-tRNA Ligase / Q000097:blood; D008970:Molecular Weight; D010766:Phosphorylation; D011817:Rabbits; D012156:Reticulocytes / Q000201:enzymology*
D015124:8-Bromo Cyclic Adenosine Monophosphate; D000604:Amino Acyl-tRNA Synthetases; D001226:Aspartate-tRNA Ligase; D005975:Glutamate-tRNA Ligase; C020659:glutaminyl-tRNA synthetase; D008250:Lysine-tRNA Ligase; D015056:1-Methyl-3-isobutylxanthine