PP60 C-SRC IN GROWTH TRANSFORMATION & DIFFERENTIATION

pp 60 c-src是pp 60 v-src的细胞同源物，pp 60 v-src是 劳斯肉瘤病毒。 蛋白质位于细胞质表面， 质膜，并被认为在细胞内的信号转导中起作用。 对有丝分裂原和其他细胞外信号包括PDGF、NGF的应答 也许还有脑脊液1 这里提出的实验利用最近 开发了重组DNA技术，并解决了两个不同的，但 pp 60 c-src的分子生物学相关问题。 的 第一组实验检查了在细胞中的结构-功能关系。 蛋白 真核病毒载体将用于获得毫克 大量的蛋白质。 纯化的pp 60 c-src应有助于 我们建议首先使用它来制备单克隆抗体， 抗体，并寻找激酶和磷酸酶，相信修改 该分子作为酪氨酸激酶的活性。 为了补充这些 我们将构建一个大型的突变c-src基因文库， 使用我们最近构建的重组鼠逆转录病毒， 以及曼贾提斯实验室的GC钳技术。 的 第二组实验旨在跟进我们最近的发现， pp 60 c-src的水平在分化过程中被调节约20倍 单核细胞。 我们设计了实验来发现 受体在完全分化的细胞中与pp 60 c-src相互作用 并观察pp 60 c-src是否也在分化中发挥作用 过程本身。

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.