SECRETORY PROTEIN SYNTHESIS DURING AGING

衰老过程中分泌蛋白的合成

• 批准号: 3118274
• 负责人: SUN-KEE KIM
• 金额: $ 6.23万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-08-01 至 1990-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3118274
• 关键词: Escherichia coli; aging; complementary DNA; eukaryote; exocrine glands; genetic transcription; genetic translation; messenger RNA; nucleic acid sequence; parotid gland; plasmids; secretion

The decline in rates of protein biosynthesis is a near universal phenomenon which takes place with age in many eukaryotic organs and tissues. The long term objective is to determine what age-related cellular changes are responsible for the decline in rates of synthesis of secretory proteins in exocrine glands, using the rat parotid gland as a model system. The immediate objective is to determine whether the changes in raters of secretory protein synthesis in rat parotid glands result from the changes in the ability of the acinar cells to produce specific messenger RNA (mRNA). The will be done by mRNA-complementary DNA (cDNA) hybridization techniques to identify and quantitate mRNA specific for alpha-amylase, the major secretory protein of the gland. DH1 strain of E. coli carrying a high copy number plasmid Puc9 with inserted salivary amylase sequence at pst 1 sites is available in this laboratory to generate amylase cDNA probe. The cDNA probe will be used to compare the age-related differences in rates of amylase gene transcription and accumulation of amylase mRNA in glands which have been depleted of stored secretory proteins stimulated discharge. Specific aims of the proposed study are to determine age-related differences in: a. the available amount of amylase mRNA in unstimulated glands. b. the lag time between stimulated secretion and accumulation of amylase mRNA, c. the rate of transcription of the amylase gene, d. the extent of amylase mRNA accumulation, e. the molecular sizes of the accumulating mRNA, f. the translational activity of total and amylase mRNA that accumulates, and g. the rate of synthesis and accumulation of amylase itself in relation to the above mRNA changes.

蛋白质生物合成速率的下降几乎是普遍的， 在许多真核生物中， 器官和组织 长期目标是确定 与年龄相关的细胞变化是导致 外分泌腺分泌蛋白的合成速率，使用 大鼠腮腺作为模型系统。 当前的目标 是为了确定分泌蛋白的变化 在大鼠腮腺中的合成是由于 腺泡细胞产生特异性信使RNA的能力 （mRNA）。 这将通过mRNA互补DNA完成 （cDNA）杂交技术来鉴定和定量mRNA 特异性α-淀粉酶，主要分泌蛋白的 腺。 DH1株E.携带高拷贝质粒Puc9的大肠杆菌 在PST1位点插入唾液淀粉酶序列的 在本实验室制备淀粉酶cDNA探针。 的cdna 将使用探针比较与年龄相关的比率差异 淀粉酶基因转录和淀粉酶积累 腺体中的mRNA已经耗尽了储存的分泌 蛋白质刺激放电。 拟议方案的具体目标 研究目的是确定以下方面与年龄有关的差异： a. 未受刺激细胞中淀粉酶mRNA的可用量 腺体 B. 刺激分泌和积累之间的滞后时间 淀粉酶mRNA， C. 淀粉酶基因的转录速率， D. 淀粉酶mRNA积累的程度， e. 积累的mRNA的分子大小， F. 总mRNA和淀粉酶mRNA翻译活性 积累，并且 G. 淀粉酶自身合成和积累速率 与上述mRNA的变化有关。