Karnitz L M, Sutor S L, Abraham R T
Department of Immunology, Mayo Clinic, Rochester, Minnesota 55905.
J Exp Med. 1994 Jun 1;179(6):1799-808. doi: 10.1084/jem.179.6.1799.
The proliferation of antigen-activated T cells is mediated by the T cell-derived growth factor, interleukin 2 (IL-2). The biochemical signaling cascades initiating IL-2-induced growth are dependent upon protein tyrosine kinase (PTK) activity. One IL-2-regulated PTK implicated in this cascade is the Src-family kinase, Fyn. Previous studies have described a physical association between Fyn and a potential downstream substrate, phosphatidylinositol 3-kinase (PI3-kinase) as well as the IL-2-dependent activation of PI3-kinase in T cells; however, the role of Fyn in IL-2-induced PI3-kinase activation remains unclear. In this report, we demonstrate that IL-2 stimulation triggers tyrosine phosphorylation of the p85 subunit of PI3-kinase in the murine T cell line, CTLL-2. Lysates prepared from growth factor-deprived and IL-2-stimulated T cells reconstituted both the binding of CTLL-2 cell-derived Fyn to and the IL-2-inducible tyrosine phosphorylation of exogenously added recombinant p85. Furthermore, overexpression of wild-type Fyn in these cells enhanced both the basal and IL-2-mediated activation of PI3-kinase. Additional studies of the Fyn-PI3-kinase interaction demonstrated that the Src homology 3 (SH3) domain of Fyn constitutes a direct binding site for the p85 subunit of PI3-kinase. These results support the notion that Fyn may be directly involved in the activation of the downstream signaling enzyme, PI3-kinase, in IL-2-stimulated T cells.
抗原激活的T细胞增殖由T细胞衍生的生长因子白细胞介素2(IL-2)介导。启动IL-2诱导生长的生化信号级联依赖于蛋白酪氨酸激酶(PTK)活性。在这一级联反应中涉及的一种受IL-2调节的PTK是Src家族激酶Fyn。先前的研究描述了Fyn与潜在的下游底物磷脂酰肌醇3激酶(PI3激酶)之间的物理关联以及T细胞中PI3激酶的IL-2依赖性激活;然而,Fyn在IL-2诱导的PI3激酶激活中的作用仍不清楚。在本报告中,我们证明IL-2刺激触发了小鼠T细胞系CTLL-2中PI3激酶p85亚基的酪氨酸磷酸化。从生长因子缺乏和IL-2刺激的T细胞制备的裂解物既重建了CTLL-2细胞衍生的Fyn与外源添加的重组p85的结合,也重建了IL-2诱导的酪氨酸磷酸化。此外,在这些细胞中野生型Fyn的过表达增强了PI3激酶的基础激活和IL-2介导的激活。对Fyn-PI3激酶相互作用的进一步研究表明,Fyn的Src同源3(SH3)结构域构成了PI3激酶p85亚基的直接结合位点。这些结果支持了Fyn可能直接参与IL-2刺激的T细胞中下游信号酶PI3激酶激活的观点。