CELL MEMBRANE SYNTHESIS IN THE CONTROL OF GROWTH

细胞膜合成对生长的控制

• 批准号: 3279976
• 负责人: LEONARD E MINDICH
• 金额: $ 25.4万
• 依托单位: PUBLIC HEALTH RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-08-01 至 1992-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3279976
• 关键词: alkaline phosphatase; bacteriophage lambda; binding proteins; cell membrane; complementary DNA; genetic manipulation; genetic transcription; genetic translation; hydrogen transport; laboratory rabbit; membrane lipids; membrane proteins; microorganism growth; mutant; nucleic acid sequence; plasmids; protein biosynthesis; protein engineering; protein structure; synthetic peptide; temperature sensitive mutant; virus envelope; virus genetics; virus protein

The assembly of the membrane of a lipid-containing bacteriophage, phi 6, will be studied as a means of elucidating the processes of membrane differentiation and translocation. Phi 6 contains a genome of three segments of double-stranded RNA inside of a ployhedral procapsid, which is in turn inside of another capsid shell. This double layered structure, called the nucleocapsid, is enveloped by a membrane that contains phospholipids and five species of protein that are specified by the phage genome. The assembly of the lipid-containing membrane on the particle is dependent upon the activity of protein P12, which is the only phage specified protein that is not a part of the virion. The assembly process has been studied in our laboratory in great detail using nonsense mutants of the phage. In addition, we have made cDNA clones of the entire genome, and we have sequenced the entire genome. We have also prepared expression vectors for both E. coli and the Pseudomonad hosts of the phage and demonstrated complementation activity of the cDNA material. We will investigate the assembly of the viral membrane by studying the interaction between protein P12 and nascent viral membrane proteins in vitro. Fusions of the membrane proteins with alkaline phosphatase will be prepared as a means of investigating whether particular parts of the proteins have targeting sequences. Physiological studies will also be performed to study the dependence of the localization of one or more of the membrane proteins on the presence of protein P12 and several other membrane proteins that may play important roles in the formation of the membrane. The formation of membranes and the transfer of membrane material is vital to many cellular processes such as organelle formation, receptor mediated responses, virus formation, and secretion of cellular materials. Understanding the processes would be of use in combating viral diseases, in ameliorating diseases due to dysfunction of hormone and metabolite receptor responses, nerve impulse transmission, cellular differentiation and proliferation.

含脂质膜的组装 将研究噬菌体 phi 6，作为阐明噬菌体 膜分化和易位的过程。 Φ6 包含三段双链RNA的基因组 多面体衣壳内部，而多面体衣壳又在另一个衣壳内部 衣壳。 这种双层结构，称为 核衣壳，被含有核衣壳的膜包裹 磷脂和五种蛋白质，由 噬菌体基因组。 含脂质膜的组装 该颗粒依赖于蛋白质 P12 的活性， 是唯一不属于病毒体一部分的噬菌体特异蛋白质。 我们的实验室对装配工艺进行了深入研究 使用噬菌体无义突变体的细节。 此外，我们还有 制作了整个基因组的 cDNA 克隆，并且我们已经测序 整个基因组。 我们还准备了表达载体 大肠杆菌和噬菌体的假单胞菌宿主 证明了 cDNA 材料的互补活性。 我们将通过以下方式研究病毒膜的组装 研究蛋白质 P12 与新生病毒之间的相互作用 体外膜蛋白。 膜蛋白的融合 用碱性磷酸酶将制备作为一种手段 研究蛋白质的特定部分是否具有 靶向序列。 还将进行生理学研究 研究一个或多个本地化的依赖性 膜蛋白对蛋白质 P12 和几种蛋白存在的影响 其他可能在细胞膜中发挥重要作用的膜蛋白 膜的形成。 膜的形成和膜的转移 材料对于许多细胞过程（例如细胞器）至关重要 形成、受体介导的反应、病毒形成，以及 细胞物质的分泌。 了解流程 可用于对抗病毒性疾病、改善 激素及代谢受体功能障碍所致疾病 反应、神经冲动传递、细胞分化和 增殖。