Zhou P, Bogacki R, McReynolds L, Howley P M
Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Mol Cell. 2000 Sep;6(3):751-6. doi: 10.1016/s1097-2765(00)00074-5.
The functional characterization of a specific gene, or its protein product, often relies on assessing the consequences of its elimination, usually accomplished by gene knockout, ribozyme, antisense, or RNA-mediated interference (RNAi) technologies. The selective degradation of cellular proteins is mediated primarily by the ubiquitin-proteasome pathway. Manipulation of the ubiquitin-dependent proteolytic machinery to eliminate specific gene products at the protein level has been previously attempted with some success in vitro; however, the in vivo efficacy of this approach has not yet been achieved. Here we report successful engineering of the substrate receptor of a major ubiquitin-proteolytic machinery to direct the degradation of otherwise stable cellular proteins both in yeast and in mammalian cells.
特定基因或其蛋白质产物的功能特性通常依赖于评估其缺失所产生的后果，这通常通过基因敲除、核酶、反义技术或RNA介导的干扰（RNAi）技术来实现。细胞蛋白质的选择性降解主要由泛素-蛋白酶体途径介导。此前曾尝试操纵泛素依赖性蛋白水解机制以在蛋白质水平消除特定基因产物，在体外取得了一定成功；然而，该方法在体内的有效性尚未实现。在此，我们报告了对一种主要泛素蛋白水解机制的底物受体进行成功改造，从而在酵母和哺乳动物细胞中直接降解原本稳定的细胞蛋白质。
