FPS/FES PROTO-ONCOGENE IN MYELOID CELL FUNCTION

C-fps/fes基因编码一种胞质酪氨酸激酶 在髓系细胞，尤其是巨噬细胞中特异表达。 这项研究的长期目标是了解 该基因的作用、作用机制及其分子基础 致癌潜力。 我们将研究c-fps/fes在信号中的可能作用。 通过确定其产物是否经历了任何生化反应来进行转导 变化(即酪氨酸磷酸化和酶的激活 活性)在髓系细胞分化和巨噬细胞激活过程中， 并通过检测重新引入的生物和生化效应 将具有生物活性的fps/fes基因导入这些细胞。生物化学 C-fps/fes动作的参数将使用温度进行解剖。 C-fps/fes的敏感和其他突变体，将被引入到 髓系细胞能够通过以下途径分化和成熟 逆转录病毒介导的基因转移。这些实验应该能澄清 C-fps/fes的生物学功能，并提供鉴定手段 增殖、分化和成熟特定的底物。我们 也会产生针对之前发现的两种抗体 C-fps/fes的底物，将用于其表征 和分子克隆。 我们将鉴定与c-fps/fes相互作用的细胞蛋白。 使用在杆状病毒中表达的纯化的c-fps/fes蛋白，以及 在细菌中表达的功能结构域，作为亲和试剂。这些 蛋白质也将被用来产生特定的单克隆和多克隆 抗体，它将与fps/fes突变体一起使用来 C-fps/fes激酶与底物的功能相互作用研究 其他细胞蛋白质。 C-fps/fes产品具有独特的生物和生化特性。 属性。对其作用机制的阐明将有助于揭示其作用机制 酪氨酸磷酸化在髓系细胞分化中的作用 和巨噬细胞的激活。因为巨噬细胞的激活起着核心作用 在对感染的免疫反应中，以及作为新的实验性疗法 依赖于使用利用酪氨酸磷酸化的髓系CSF 白血病细胞分化的途径、机制 这些研究发现具有广泛的潜在临床意义。

The c-fps/fes gene encodes a cytoplasmic tyrosine kinase that is specifically expressed in myeloid cells, in particular in macrophages. The long-term objective of this study is to understand the biological role and mechanism of action of this gene, and the molecular basis of its oncogenic potential. We will investigate the possible role of c-fps/fes in signal transduction by determining if its product undergoes any biochemical changes (i.e. tyrosine phosphorylation and activation of enzymatic activity) during myeloid cell differentiation and macrophage activation, and by examining the biological and biochemical effects of reintroducing biologically active fps/fes genes into these cells. The biochemical parameters of c-fps/fes action will be dissected using temperature sensitive and other mutants of c-fps/fes, which will be introduced into myeloid cells capable of undergoing differentiation and maturation by retroviral mediated gene transfer. These experiments should clarify the biological functions of c-fps/fes and provide the means to identify proliferation-, differentiation-, and maturation-specific substrates. We will also generate antibodies against two previously identified substrates of c-fps/fes, which will be used for their characterization and molecular cloning. We will identify cellular proteins that interact with c-fps/fes kinase using purified c-fps/fes protein expressed in baculovirus, and functional domains expressed in bacteria, as affinity reagents. These proteins will also be used to generate specific monoclonal and polyclonal antibodies, which will be used in conjunction with fps/fes mutants to study functional interactions of c-fps/fes kinase with substrates and other cellular proteins. The c-fps/fes product has unique biological and biochemical properties. The elucidation of its mechanism of action will shed light on the role of tyrosine phosphorylation in myeloid cell differentiation and macrophage activation. As macrophage activation plays a central role in the immune response to infections, and as new experimental therapies rely on the use of myeloid CSFs that utilize tyrosine phosphorylation pathways to achieve differentiation of leukemic cells, the mechanisms uncovered in these studies have broad potential clinical implications.