INDUCED SELECTIVE DEGRADATION OF CELLULAR PROTEINS

描述：(改编自申请者摘要)：本报告的总体目标 项目是开发和利用一种系统，借此特定的细胞靶点 一种或多种蛋白质可以根据指令被蛋白酶体破坏。建议数 工作将促进生物技术的发展，通过这种生物技术， 蛋白质表达、合成和降解的重要元素可能是 有效地解决了这些问题。研究人员建议利用蛋白质模块 正常情况下直接调节鸟氨酸脱羧酶(ODC)的降解，通过 把它们送到选定的目标。两个关键组件是C ODC末端(CODC)和抗酶N末端的一部分(NAZ)。他们 将增强的绿色荧光用作方便检测的目标 蛋白质(绿色荧光蛋白)。他们将通过使用一个 包含FKBP12和FKBP12的两种融合蛋白的条件性结合方法 FRB结构域，这取决于药物雷帕霉素。无论是体外还是体内 将测试(在培养细胞中)的降解情况。首先，他们将确定 FRB-EGFP-CODC在绑定到NAZ-FKBP12时是否被降解。第二，他们会 确定诱导性翻译移码是否可以用来调节 这一过程的活动。第三，他们将确定CODC是否也可以 被用作条件降级信号。 建议的商业应用：不可用

DESCRIPTION: (Adapted from applicant's abstract): The overall aim of this project is to develop and exploit a system whereby a specific cellular target protein or proteins can be destroyed by the proteasome on command. The proposed work will facilitate the development of a biotechnology through which both important elements of protein expression, synthesis, and degradation can be addressed effectively. The investigators propose to utilize protein modules that normally direct regulated degradation of ornithine decarboxylase (ODC), by delivering them to targets of choice. The two critical components are the C terminus of ODC (CODC) and a portion of the N terminus of antizyme (NAZ). They will use as a conveniently assayed target the enhanced green fluorescent protein (EGFP). They will test the feasibility of this approach by using a means of conditional association of two fusion proteins, containing FKBP12 and FRB domains, that depends on the drug rapamycin. Both in vitro and in vivo degradation (in cultured cells) will be tested. First, they will determine whether FRB-EGFP-CODC is degraded when bound to NAZ-FKBP12. Second, they will determine whether induced translational frameshifting can be used to modulate the activity of this process. Third, they will determine whether CODC can also be used as a conditional degradation signal. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE