INTERMEDIARY METABOLISM AND A NOVEL RNA BINDING PROTEIN

中间代谢和新型 RNA 结合蛋白

• 批准号: 2691386
• 负责人: Clay F. Semenkovich
• 金额: $ 19.75万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-29 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2691386
• 关键词: RNA binding protein; adipose tissue; cell growth regulation; cell line; diabetes mellitus; disease /disorder model; enzyme activity; enzyme induction /repression; fatty acid biosynthesis; fatty acid synthase; gene expression; laboratory mouse; molecular cloning; protein binding; ribozymes; transfection

Fatty acid synthase (FAS) is an enzyme critical for de novo fatty acid synthesis. Glucose regulates human FAS expression by mediating FAS mRNA stability. Glucose also induces the binding of a 178 kD protein to the FAS message, suggesting that this protein links intermediary metabolism and fatty acid biosynthesis. This RNA binding protein has been purified and partially sequenced. Its major binding element has been identified. The studies in this application will test the hypothesis that this glucose-inducible protein regulates de novo fatty acid synthesis by mediating FAS gene expression. The specific aims are: 1-To clone and express the 178 kD FAS mRNA binding protein. 2-To determine the binding activity of the 178 kD protein in different lipogenic tissues in the setting of experimental diabetes in mice. 3-To demonstrate the involvement of the 178 kD binding element in message stability. 4-To inactivate the 178 kD protein in HepG2 cells and determine effects on lipoprotein secretion and cell growth. 5-To overexpress the 178 kD protein in HepG2 cells and determine effects on lipoprotein secretion and cell growth. We have identified what appears to be a novel RNA binding protein. Characterizing this RNA binding protein could provide the basis for innovative therapies aimed at human disorders associated with abnormal fatty acid and glucose metabolism. These disorders include obesity, hyperlipidemia, non-insulin-dependent diabetes mellitus and even malignancies.

脂肪酸合成酶（FAS）是脂肪酸从头合成的关键酶 合成. 葡萄糖介导FAS mRNA调节人FAS表达 稳定 葡萄糖还诱导178 kD蛋白与 FAS信息，表明这种蛋白质连接中间代谢 和脂肪酸生物合成。 该RNA结合蛋白已被纯化 并部分测序。 它的主要结合元素已经确定。 本申请中的研究将检验这一假设， 葡萄糖诱导蛋白调节从头脂肪酸合成， 介导FAS基因表达。 具体目标是：1-克隆和 表达178 kD FAS mRNA结合蛋白。2-确定约束力 178 kD蛋白在不同脂肪生成组织中的活性 小鼠实验性糖尿病的设置。3-证明 178 kD结合元件参与信息稳定性。4-至 在HepG 2细胞中表达178 kD蛋白并测定其对HepG 2细胞增殖的影响 脂蛋白分泌和细胞生长。 5-过度表达178 kD HepG 2细胞中的蛋白质，并确定对脂蛋白分泌的影响 和细胞生长。 我们发现了一种新的RNA结合蛋白。 这种RNA结合蛋白的特性可以为 创新疗法，针对与异常 脂肪酸和葡萄糖代谢。这些疾病包括肥胖症， 高脂血症、非胰岛素依赖型糖尿病，甚至 恶性肿瘤。