PP60 C-SRC IN GROWTH TRANSFORMATION & DIFFERENTIATION

Pp60c-src是Ppp60v-src的细胞同源物，pp60v-src是 劳斯肉瘤病毒。该蛋白位于细胞质面。 质膜，并被认为在信号转导中起作用 对有丝分裂原和包括PDGF、NGF在内的其他细胞外信号的反应 或许还有脑脊液-1。这里提出的实验利用了最近的 开发了重组DNA技术，并解决了两个不同的问题，但 与pp60c-src相关的分子生物学问题。这个 第一组实验考察了结构与功能之间的关系 蛋白。将使用真核病毒载体获得毫克 蛋白质的数量。纯化的pp60c-src应该有助于 实验次数-我们建议首先使用它来制备克隆 抗体，并寻找被认为可以修饰的激酶和磷酸酶 分子作为酪氨酸酶的活性。为了补充这些内容 生化研究我们将构建一个突变c-src基因的大型文库。 使用我们最近构建的重组小鼠逆转录病毒 以及曼贾提斯实验室的所谓GC钳技术。这个 第二组实验是为了跟进我们最近的发现 在分化过程中，pp60c-src的水平被调节了约20倍。 单核细胞。我们设计了实验，以发现特定的 在完全分化的细胞中，受体与pp60c-src相互作用 来看看pp60c-src是否也在分化中起作用。 进程本身。

pp60c-src is the cellular homolog of pp60v-src, the transforming protein of Rous Sarcoma Virus. The protein is located on the cytoplasmic face of the plasma membrane and is believed to function in signal transduction in response to mitogens and other extracellular signals including PDGF, NGF and, perhaps, CSF-1. The experiments proposed here make use of recently developed recombinant DNA techniques and address two different, but related, questions concerning the molecular biology of pp60c-src. The first set of experiments examines structure-function relationships in the protein. Eukaryotic viral vectors will be used to obtain milligram quantities of the protein. The purified pp60c-src should facilitate a number of experiments - we propose using it first to prepare monoclonal antibodies and to search for kinases and phosphatases believed to modify the activity of the molecule as a tyrosine kinase. To complement these biochemical studies we will construct a large library of mutant c-src genes using a recombinant murine retrovirus which we have recently constructed and the so-called GC clamp technique from the Manjatis laboratory. The second set of experiments are designed to follow up our recent finding that the level of pp60c-src is modulated some 20-fold during the differentiation of monocytes. We have designed experiments to discover if specific receptors are interacting with pp60c-src in the fully differentiated cells and to see if pp60c-src might also play a role in the differentiation process itself.