MECHANICS AND FUNCTIONS OF THE N-END RULE PATHWAY

N 端规则路径的机制和功能

• 批准号: 6150610
• 负责人: ALEXANDER J VARSHAVSKY
• 金额: $ 42.04万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-02-01 至 2003-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6150610
• 关键词: Saccharomyces cerevisiae; fungal genetics; genetic techniques; microorganism metabolism; mitochondria; mutant; posttranslational modifications; proteasome; protein degradation; protein structure function; proteolysis; subtraction hybridization; ubiquitin

DESCRIPTION: Ubiquitin is a 76-residue protein that exists in cells either free or conjugated to many other proteins. Selective degradation of proteins by the ubiquitin/proteasome-dependent pathways plays a role in a multitude of biological processes, including cell growth and differentiation, signal transduction and responses to stress. One ubiquitin/proteasome-dependent proteolytic system is the N-end rule pathway, identified by the laboratory in 1986. The N-end rule relates the in vivo half-life of a protein to the identity of its N-terminal residue. The N-end rule pathway is present in all organisms examined, from mammals to bacteria. Studies supported by the present grant ("Mechanics and Functions of the N-end Rule Pathway", DK39520), currently in its 11th year, have yielded significant insights into the N-end rule's functions and mechanisms, as described in the Progress Report. The objective of the research described in this renewal application is to advance the understanding of the N-end rule pathway and related aspects of the ubiquitin system. Specific aims are: 1) Regulation of peptide import by the N-end rule pathway: biochemical and genetic studies; 2) A fusion-based screen for physiological substrates of the N-end rule pathway in S. cerevisiae; 3) Isolation and analysis of regulators of the N-end rule pathway; 4) Screens for mutants whose viability requires the presence of the N-end r; 5) The use of suppression subtractive hybridization to identify new functions of the N-end rule pathway; 6) Analysis of a mitochondrial function of the Ntalp N-terminal amidase in S. cerevisiae; 7) A biochemico-genetic approach to identification of physiological N-end rule substrates; 8) A "double-headed" inhibitor of the N-end rule pathway; 9) The "two-ubiquitin" technique and its application to the problem of cotranslational proteolysis; and 10) Biochemical and genetic dissection of the UBRI-encoded Nrecognin and Ubr2p, a recently identified homolog of Ubrlp in S. cerevisiae.

描述：泛素是一种 76 个残基的蛋白质，存在于细胞中 游离或与许多其他蛋白质结合。 选择性降解 泛素/蛋白酶体依赖性途径的蛋白质在 多种生物过程，包括细胞生长和 分化、信号转导和应激反应。 一 泛素/蛋白酶体依赖性蛋白水解系统是N端规则途径， 1986年由实验室鉴定。N端规则与体内 蛋白质的半衰期与其 N 末端残基的身份有关。 N端 规则途径存在于所有被检查的生物体中，从哺乳动物到细菌。 目前资助的研究（“力学和功能 N端规则途径”，DK39520），目前已进入第11个年头，已产生 对 N 端规则的功能和机制的重要见解，如 进展报告中描述。 描述的研究目的 本次更新应用是为了加深对N端的理解 泛素系统的规则途径和相关方面。 具体目标 是： 1) N端规则途径对肽输入的调节： 生化和遗传学研究； 2）基于融合的生理筛选 酿酒酵母 N 端规则途径的底物； 3）隔离和 N端规则通路的调节因子分析； 4) 筛选突变体 其生存能力需要 N 端 r 的存在； 5）使用 抑制消减杂交鉴定N端新功能 规则路径； 6) Ntalp线粒体功能分析 酿酒酵母中的 N 末端酰胺酶； 7) 生物化学遗传学方法 生理N端规则底物的鉴定； 8）“双头” N端规则途径的抑制剂； 9）“双泛素”技术和 其在共翻译蛋白水解问题中的应用；和 10) UBRI 编码的 Nrecognin 和 Ubr2p 的生化和遗传解剖， 最近在酿酒酵母中发现了 Ubrlp 的同源物。