DIFFERENTIAL REGULATION OF GLUCOSE TRANSPORTER ISOFORMS

葡萄糖转运蛋白异构体的差异调节

• 批准号: 3464793
• 负责人: MARTYN K WHITE
• 金额: $ 8.1万
• 依托单位: EAST CAROLINA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1997-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3464793
• 关键词: animal tissue; chickens; chimeric proteins; gene deletion mutation; genetic regulation; genetic transcription; glucose transport; immunoelectron microscopy; membrane proteins; nucleic acid sequence; oncogenes; protein biosynthesis; protein degradation; protein structure function; regulatory gene; transport proteins

The rate of glucose uptake into vertebrate cells is regulated by oncogenic transformation and by many other factors including mitogen, hormones and glucose starvation. It has been shown that regulation of glucose transporters can occur at different levels including transcription, mRNA stability, protein translocation and protein turnover. Glucose transporters provide a way of studying cellular regulation at these levels as well as the relationship of growth control to carbohydrate metabolism. In addition there are five transporter isoforms which are regulated differently, thus providing a useful model system for understanding how isoforms are regulated according to the metabolic needs of the cell. We are interested in chicken embryo fibroblasts since they express at least two glucose transporter isoforms and these are differentially regulated (type 1 is regulated by protein turnover and type 3 by the level of its mRNA). The goals of this proposal are to understand the molecular basis of this differential regulation. AIM 1: We propose to complete the cloning and sequencing of the cDNAs of glucose transporter isoforms expressed by chicken embryo fibroblasts. AIM 2: These will be used to measure the levels of the mRNAs for each isoform under various physiological conditions. The rate of transcription for each isoform and stability of its mRNA will also be measured. AIM 3: The molecular basis for the differential transcriptional control of the transporter isoforms will be determined by isolating genomic clones and analyzing their promoter regions. AIM 4: The control of transporters at the protein level will be investigated by generating specific antisera to each isoform. These will be used to quantitate the level of each isoform under various physiological conditions and to measure their rates of biosynthesis and turnover. It has already been established that the type 1 isoform is regulated at the level of turnover. AIM 5: Cellular and biochemical analysis of the molecular basis of this turnover control will be undertaken. This study will provide basic information about the regulation of different isoforms and about several levels of cellular control.

葡萄糖摄取对脊椎动物细胞的摄取速率受致癌性调节 转化以及许多其他因素，包括有丝分裂原，激素和 葡萄糖饥饿。 已经表明葡萄糖的调节 转运蛋白可以在不同级别发生，包括转录，mRNA 稳定性，蛋白质易位和蛋白质更新。 葡萄糖 运输者提供了一种研究这些水平的细胞调节的方法 以及生长控制与碳水化合物代谢的关系。 另外，有五个受调节的转运蛋白同工型 以不同的方式，因此提供了一个有用的模型系统来理解如何 根据细胞的代谢需求调节同工型。 我们 对鸡肉胚胎成纤维细胞感兴趣，因为它们至少表达 两个葡萄糖转运蛋白同工型，它们受到差异调节 （1型由蛋白质周转和3型调节。 mRNA）。 该提议的目标是了解 这种差异调节。 目标1：我们建议完成的克隆和测序 由鸡胚胎成纤维细胞表达的葡萄糖转运蛋白同工型。 目的 2：这些将用于测量每种同工型的mRNA水平 在各种生理条件下。 每个转录率 也将测量其mRNA的同工型和稳定性。 目标3： 分子基础，用于差分转录对照 转运蛋白同工型将通过分离基因组克隆和 分析其发起人区域。 目标4：转运蛋白的控制 蛋白质水平将通过产生特定的抗血清来研究 每个同工型。 这些将用于定量每个同工型的水平 在各种生理条件下，衡量其率 生物合成和周转。 已经确定了类型 1同工型在营业额的水平下进行调节。 目标5：细胞和 该离职控制的分子基础的生化分析将 进行。 这项研究将提供有关 调节不同同工型和大约几个细胞 控制。