BIOCHEMISTRY OF THE CANCER CELL

本提案中概述的研究目标是定义 转化基因产物(如pp60-v-src和p21-v-Ha-ras)的作用， 血清生长因子或促肿瘤佛波酯在生长控制和治疗中的作用 细胞转化。转化蛋白与蛋白质的相互作用 将对细胞膜进行表征，并尝试确定 任何细胞膜组件都是膜结合所必需的 使用纯化的蛋白质组分和合成的脂囊泡 重建实验。我们将研究细胞酶，它可能是 有助于引起细胞所显示的表型变化 对这些转化蛋白或肿瘤促进剂的反应。举例 这类细胞酶包括丝氨酸特异性核糖体蛋白S6。 激酶、参与调节环磷酸腺苷水平的酶和酶 参与第二信使二甘油和甘油的产生 三磷酸肌醇。如果可行，这些细胞酶将被使用 产生多克隆或单克隆抗体作为辅助 功能鉴定及其在获得cdna或基因组中的应用 这些蛋白质的分子克隆。我们把静止的细胞比作 刺激增殖的细胞合成特异性 蛋白质、蛋白质磷酸化模式与细胞表达 基因。这些研究应该会提供关于 导致细胞增殖和恶性的生化途径 转型。 我们将利用1)多种技术进行蛋白质纯化；2) C DNA文库的分子克隆；3)多克隆和单克隆 抗体；4)病毒转化突变体。

The objectives of the research outlined in this proposal are to define the role of transforming gene products (such as pp60-v-src and p21-v-Ha-ras), serum growth factors or tumor promoting phorbol ester in growth control and cell transformation. The interactions of the transforming proteins with cellular membranes will be characterized and attempts made to determine if any cellular membrane components are required for membrane association by using purified protein components and synthetic lipid vesicles in reconstitution experiments. We shall study cellular enzymes that may be instrumental in causing the phenotypic changes displayed by cells in response to these transforming proteins or tumor promoters. Examples of such cellular enzymes include the serine-specific ribosomal protein S6 kinase, enzymes involved in the regulation of cyclic AMP levels and enzymes involved in the generation of the second messengers diacylglycerol and inositol trisphosphate. When feasible, these cellular enzymes will be used to generate polyclonal or monoclonal antibody as an additional aid in functional characterization and for use in obtaining cDNA or genomic molecular clones for these proteins. We shall compare quiescent cells to cells stimulated to proliferate with regard to synthesis of specific proteins, protein phosphorylation patterns and expression of cellular genes. These studies should provide new insights concerning the biochemical pathways that lead to cellular proliferation and malignant transformation. We shall utilize 1) multiple techniques for protein purification; 2) molecular cloning of cDNA libraries; 3) polyclonal and monoclonal antibodies; 4) viral transformation mutants.