Differential regulation of oxidative and osmotic stress induced Syk activation by both autophosphorylation and SH2 domains.
Syk, a nonreceptor protein-tyrosine kinase, is activated by both oxidative and osmotic stress and plays different roles in the transduction of stress signals. In this study, the regulation of oxidative and osmotic stress induced Syk activation was investigated utilizing Syk-negative DT40 cells, expressing various Syk mutants. Phosphorylation of Y518Y519 was demonstrated to be required for both oxidative and osmotic stress induced Syk activation. Syk activation by these two types of stress stimuli was a combination of both autophosphorylation and the activities of additional tyrosine kinases. Oxidative stress induced Syk tyrosine phosphorylation was almost completely attributed to autophosphorylation, whereas other tyrosine kinases were largely responsible for osmotic stress induced Syk tyrosine phosphorylation. Moreover, the Src homology 2 (SH2) domains of Syk differentially regulated Syk activation. Both mSH2(N) Syk and mSH2(C) Syk, in which the phosphotyrosine-dependent binding motif within the SH2 domains contained point mutations, showed a significantly higher activity than that observed in wild-type Syk, following osmotic stress treatment. In comparison, in response to oxidative stress, only mSH2(N) Syk demonstrated a stronger activation than wild-type Syk. Therefore, differential activation and regulation of Syk may give an insight into the distinctive functions of Syk in oxidative and osmotic stress signaling.
Qin
S
S
Department of Biochemistry, Kobe University School of Medicine, Japan.
Kurosaki
T
T
Yamamura
H
H
D000818:Animals; D002460:Cell Line; D002645:Chickens; D004789:Enzyme Activation / Q000235:genetics; D004792:Enzyme Precursors / Q000235:genetics / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins; D009997:Osmotic Pressure; D018384:Oxidative Stress / Q000235:genetics; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000235:genetics; D013552:Swine; D000072377:Syk Kinase; D014162:Transfection; D014443:Tyrosine / Q000378:metabolism; D018909:src Homology Domains / Q000235:genetics / Q000502:physiology*
D004792:Enzyme Precursors; D047908:Intracellular Signaling Peptides and Proteins; D010750:Phosphoproteins; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D000072377:Syk Kinase