Phosphorylation of ribosomal protein S19 at Ser59 by CaM kinase I alpha.
In order to examine the possible involvements of Ca(2+)/calmodulin-dependent protein kinases (CaM kinases) in the regulation of ribosomal functions, we tested the phosphorylation of rat ribosomal protein S19 (RPS19) by various CaM kinases in vitro. We found that CaM kinase Ialpha, but not CaM kinase Ibeta1, Ibeta2, II, or IV, robustly phosphorylated RPS19. From the consensus phosphorylation site sequence, Ser59, Ser90, and Thr124 were likely to be phosphorylated; therefore, we mutated each amino acid to alanine and found that the mutation of Ser59 to alanine strongly attenuated phosphorylation by CaM kinase Ialpha, suggesting that Ser59 was a major phosphorylation site. Furthermore, we produced a specific antibody against RPS19 phosphorylated at Ser59, and found that Ser59 was phosphorylated both in GT1-7 cells and rat brain. Phosphorylation of RPS19 in GT1-7 cells was inhibited by KN93, an inhibitor of CaM kinases. Immunoblot analysis after subcellular fractionation of rat brain demonstrated that phosphorylated RPS19 was present in 80S ribosomes. Phosphorylation of RPS19 by CaM kinase Ialpha augmented the interaction of RPS19 with the previously identified S19 binding protein. These results suggest that CaM kinase Ialpha regulates the functions of RPS19 through phosphorylation of Ser59.
Maeda
Noriko
N
Department of Biochemistry, University of the Ryukyus, Okinawa, Japan.
Toku
Seikichi
S
Naito
Yasuhito
Y
Nishiura
Hiroshi
H
Tanaka
Tatsuo
T
Yamamoto
Hideyuki
H
D000818:Animals; D054729:Calcium-Calmodulin-Dependent Protein Kinase Type 1 / Q000378:metabolism*; D002461:Cell Line, Transformed; D029721:Drosophila Proteins / Q000378:metabolism*; D010766:Phosphorylation; D021122:Protein Subunits / Q000378:metabolism; D051381:Rats; D012269:Ribosomal Proteins / Q000378:metabolism*; D012694:Serine / Q000378:metabolism*
D029721:Drosophila Proteins; D021122:Protein Subunits; D012269:Ribosomal Proteins; C040343:ribosomal protein S19; D012694:Serine; D054729:Calcium-Calmodulin-Dependent Protein Kinase Type 1; C517599:Camk1 protein, rat