PP60 C-SRC IN GROWTH TRANSFORMATION & DIFFERENTIATION

增长转型中的 PP60 C-SRC

• 批准号: 3186168
• 负责人: THOMAS M ROBERTS
• 金额: $ 11.06万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-01-01 至 1991-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3186168
• 关键词: Adenoviridae; Rous sarcoma virus; X ray crystallography; affinity chromatography; antibody formation; autoradiography; cell differentiation; cell growth regulation; cell transformation; enzyme linked immunosorbent assay; fluorescence microscopy; gel electrophoresis; gene expression; gene mutation; genetic library; genetic manipulation; immunofluorescence technique; immunoprecipitation; laboratory mouse; molecular biology; nucleic acid hybridization; oncogenes; phosphates; plasmids; point mutation; protein biosynthesis; protein structure; protein tyrosine kinase; virus protein

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.

PP60C-SRC是PP60V-SRC的细胞同源物，是转化的蛋白 Rous肉瘤病毒。 蛋白质位于胞质面上 质膜，据信在信号转导中起作用 对有丝分裂剂和其他细胞外信号（包括PDGF，NGF）的反应 也许是CSF-1。 这里提出的实验最近使用 开发了重组DNA技术，并解决了两种不同，但 相关，有关PP60C-SRC分子生物学的问题。 这 第一组实验检查了结构 - 功能关系 蛋白质。 真核病毒载体将用于获得毫克 蛋白质的数量。 纯化的PP60C-SRC应促进 实验数 - 我们首先建议使用它来制备单克隆 抗体并搜索被认为可以修饰的激酶和磷酸酶 该分子作为酪氨酸激酶的活性。 补充这些 生化研究我们将构建一个大型突变C-SRC基因的库 使用我们最近构建的重组鼠逆转录病毒 以及Manjatis实验室的所谓GC夹具技术。 这 第二组实验旨在跟进我们最近的发现 在分化过程中调制了PP60C-SRC的水平约20倍 单核细胞。 我们设计了实验，以发现是否具体 受体正在与完全分化的细胞中的PP60C-SRC相互作用 看看PP60C-SRC是否也可能在分化中发挥作用 处理本身。