title: Phosphorylation of SET protein at Ser171 by protein kinase D2 diminishes its inhibitory effect on protein phosphatase 2A.
abstract: We previously reported that protein kinase D2 (PKD2) in T cells is promptly activated after T-cell receptor (TCR) stimulation and involved in the activation of interleukin-2 promoter and T cell death, and that one of its candidate substrate is SET protein, a natural inhibitor for protein phosphatase 2A (PP2A). In this study, we investigated the target amino acid residues of SET phosphorylated by PKD2 and the effects of phosphorylation of SET on PP2A phosphatase activity. In vitro kinase assay using various recombinant SET mutants having Ser/Thr to Ala substitutions revealed that Ser171 of SET is one of the sites phosphorylated by PKD2. Recombinant SET with phosphorylation-mimic Ser171 to Glu substitution reduced its inhibitory effects on PP2A phosphatase activity compared with Ser171 to Ala substituted or wild-type SET. In addition, knockdown of PKD2 in Jurkat cells by RNAi or treatment of human CD4(+) T cell clone with the PKD2 inhibitor Gö6976 resulted in reduced PP2A activity after TCR-stimulation judged from phosphorylation status of Tyr307 of the catalytic subunit of PP2A. These results suggest that PKD2 is involved in the regulation of PP2A activity in activated T cells through phosphorylation of Ser171 of SET.
authors.affiliation: Department of Immunogenetics, Graduate School of Medical Sciences, Kumamoto University, Honjo, Kumamoto, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Irie
authors.forename: Kumiko
authors.initials: K
authors.lastname: Harada
authors.forename: Norie
authors.initials: N
authors.lastname: Araki
authors.forename: Yasuharu
authors.initials: Y
authors.lastname: Nishimura
mesh_terms: D019943:Amino Acid Substitution; D015153:Blotting, Western; D004268:DNA-Binding Proteins; D015180:Electrophoresis, Gel, Two-Dimensional; D056488:Histone Chaperones / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D013058:Mass Spectrometry; D010766:Phosphorylation; D000091283:Protein Kinase D2; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D054648:Protein Phosphatase 2 / Q000037:antagonists & inhibitors*; D034622:RNA Interference; D012694:Serine / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D056488:Histone Chaperones; D000091283:Protein Kinase D2; C513473:SET protein, human; D014157:Transcription Factors; D012694:Serine; D011494:Protein Kinases; D054648:Protein Phosphatase 2