FPS/FES PROTO-ONCOGENE IN MYELOID CELL FUNCTION

c-fps/fes基因编码一种胞质酪氨酸激酶， 在骨髓细胞中特异性表达，特别是在巨噬细胞中。 这项研究的长期目标是了解生物学 该基因的作用和作用机制，及其分子基础， 致癌潜力 我们将研究c-fps/fes在信号转导中的可能作用 通过确定其产物是否经历任何生物化学过程来进行转导 改变（即酪氨酸磷酸化和酶的激活） 活性）在骨髓细胞分化和巨噬细胞活化期间， 通过研究重新引入的生物学和生物化学效应， 将具有生物活性的FPS/FES基因导入这些细胞中。 生化 将使用温度来剖析C-FPS/FES动作的参数 敏感和其他突变体的c-fps/fes，这将被引入到 能够通过以下方式进行分化和成熟的骨髓细胞： 逆转录病毒介导的基因转移 这些实验应该澄清 c-fps/fes的生物学功能，并提供鉴定 增殖、分化和成熟特异性底物。 我们 也会产生针对两种先前鉴定的 c-fps/fes的衬底，这将用于它们的表征 和分子克隆。 我们将鉴定与c-fps/fes相互作用的细胞蛋白质 使用在杆状病毒中表达的纯化的c-fps/fes蛋白的激酶，和 在细菌中表达的功能域，作为亲和试剂。 这些 蛋白质也将用于产生特异性单克隆和多克隆抗体 抗体，将与fps/fes突变体结合使用， 研究c-fps/fes激酶与底物功能相互作用， 其他细胞蛋白质。 c-fps/fes产品具有独特的生物和生化特性， 特性. 阐明其作用机制将有助于阐明 酪氨酸磷酸化在骨髓细胞分化中的作用 和巨噬细胞活化。 由于巨噬细胞活化在 在感染的免疫反应中， 依赖于利用酪氨酸磷酸化的髓样CSF的使用 途径，以实现白血病细胞的分化，机制 在这些研究中发现的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.