PROTEINS IN LEAD-INDUCED NUCLEAR INCLUSION BODIES

铅诱发的核包涵体中的蛋白质

• 批准号: 3249746
• 负责人: KEITH R SHELTON
• 金额: $ 13.14万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-08-01 至 1990-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3249746
• 关键词: autoradiography; electrofocusing; electron microscopy; environmental toxicology; gel electrophoresis; gene expression; genes; genetic regulation; inclusion body; laboratory mouse; laboratory rat; lead poisoning; molecular cloning; monoclonal antibody; nucleic acid hybridization; nucleic acid sequence; nucleoproteins; phase contrast microscopy; protein biosynthesis; protein metabolism; protein structure; radionuclides; renal cortex; tissue /cell culture

Lead poisoning induces nuclear bodies, composed of lead and protein, in many animals including man. These bodies are prominent in renal tubular lining cells. There are well known clinical effects of lead poisoning, but neither the function nor the effect of the nuclear inclusion bodies is known. They may reflect a regulated process which decreases the level of soluble lead. Their formation is blocked in cell culture by inhibitors of RNA and protein synthesis, suggesting that inducible proteins are involved. The inclusion bodies contain a prominent protein which has not been detected elsewhere, suggesting that it is either induced or sequestered by lead. In order to understand inclusion body formation and function, it is necessary to relate the metabolism of these proteins to lead exposure and to study the interactions of the proteins with lead. The chemical properties of the inclusion bodies impose severe restraints on their study. A cell culture system will be used in correlating synthesis of inclusion body proteins with lead exposure. Further, the synthesis of the cognate mRNA's will be determined by an in vitro translation assay in order to confirm induction by lead. The tendency of these proteins to bind lead will be studied by exposure of purified, bound protein to soluble radioactive lead. Monoclonal antibodies will be prepared and used for assaying and purifying the proteins and in characterizing their intracellular location(s). Complementary DNA clones will be isolated in order that regulation of gene activity can be studied quantitatively in various cells and under various stimuli such as lead, other metals, and other traumatic effects such as heat. The effects of lead on cells have been difficult to study at the molecular level. However, the application of recently developed procedures to a study of the long recognized nuclear inclusion bodies provides a unique opportunity to study this problem.

铅中毒导致脑组织中由铅和蛋白质组成的核小体 包括人类在内的许多动物。这些小体突出于肾小管。 衬里单元格。铅中毒的临床影响是众所周知的，但 核包涵体的功能和效果都不是 为人所知。它们可能反映了一个受监管的过程，该过程降低了 可溶铅。它们的形成在细胞培养中被抑制物阻止 RNA和蛋白质合成，表明可诱导的蛋白质是 牵涉其中。包涵体含有一种突出的蛋白质，它没有 在其他地方被检测到，表明它要么是被诱导的，要么是 被铅隔绝了。为了了解包涵体的形成和 功能，有必要将这些蛋白质的代谢与 铅暴露，并研究蛋白质与铅的相互作用。这个 包涵体的化学性质严重限制了 他们的书房。细胞培养系统将用于相关合成 有铅暴露的包涵体蛋白。此外，合成了一种新的化合物 同源信使核糖核酸将通过体外翻译试验确定 命令确认铅诱导。这些蛋白质结合的趋势 铅的研究将通过将纯化的结合蛋白暴露在可溶的环境中来进行 放射性铅。将制备单克隆抗体并用于 蛋白质的分析、纯化及其性质的研究 细胞内定位(S)。互补的DNA克隆将在 为了能够定量地研究基因活性的调节， 各种电池和在各种刺激下，如铅、其他金属和 其他创伤性影响，如高温。铅对细胞的影响有 很难在分子水平上进行研究。但是，该应用程序 最近开发的程序来研究长期被认识到的核 包涵体为研究这一问题提供了独特的机会。