Increase in the specific activity of p50 csk in proliferating T cells correlates with decreased specific activity of p56 lck and p59 fyn and reduced phosphorylation of CD3 subunits

Abstract

Depending on their prior antigen recognition history, mature T cells respond with different functional outcomes to T cell receptor (TCR) stimulation. These functional outcomes include proliferation, anergy and cell death. The biochemical basis underlying differential responses by mature T cells at different stages of their developmental pathway to TCR stimulation remains to be determined. We have previously shown that proliferating but not naive T cells were susceptible to apoptosis after TCR stimulation and that the tyrosine phosphorylation of TCRζ, CD3γ, and CD3ϵ in proliferating T cells was decreased after TCR stimulation. In this study, we determined whether differences in phosphorylation between naive and proliferating T cells were due to altered regulation of p56 lck (Lck) or p59 fyn (Fyn) by their positive or negative regulators, CD45 or p50 csk (Csk), respectively. We found that Lck was expressed at the same level and had the same phosphotyrosine content in naive and proliferating T cells. However, its autophosphorylation activity was lower in proliferating cells, corresponding to a 2-fold decrease in its specific kinase activity. Similarly, the specific kinase activity of Fyn was also decreased by about 2-fold in proliferating T cells. In contrast, although Csk was expressed at the same level in both cell types its specific kinase activity was increased by 6-fold in proliferating T cells. The tyrosine phosphatase CD45, a positive regulator of src-family kinases, was overexpressed by 3- to 6-fold in proliferating cells. However, the specific activity of CD45 in naive and proliferating T cells was the same. Therefore, although the protein expression level of CD45 was increased in proliferating T cells it only partially compensated for the hyperactivity of Csk resulting in a 2-fold reduction in the specific activity of Lck and Fyn in proliferating T cells.