Chai Bao-Feng, Wang Wei, Liang Ai-Hua
Institute of Biotechnology, Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi Univesity, Taiyuan, PR China.
Res Microbiol. 2006 Apr;157(3):235-40. doi: 10.1016/j.resmic.2005.08.001. Epub 2005 Sep 19.
Class II polypeptide release factor eRF3 stimulates translation termination in a GTP-dependent manner. Recent data show that eRF3 is also involved in cell cycle regulation apart from its function in translation. Here we show that recombinant Eo-eRF3 from the ciliate Euplotes octocarinatus expressed in Escherichia coli can be tested for its ribosome-dependent GTPase activity using an in vitro system. Moreover, polyclonal antibodies against recombinant Eo-eRF3 were used to determine the localization of Eo-eRF3 using immunofluorescence and immunoelectron microscopy, which showed that the Eo-eRF3 is distributed not only around the macronucleus, but also in basal bodies in the cortex of E. octocarinatus cells. The results suggest that Eo-eRF3 may be involved in cytoskeleton organization in addition to its function in translation termination.
II类多肽释放因子eRF3以GTP依赖的方式刺激翻译终止。最近的数据表明，eRF3除了在翻译中发挥作用外，还参与细胞周期调控。在这里，我们表明，在大肠杆菌中表达的来自八肋游仆虫的重组Eo-eRF3可以使用体外系统检测其核糖体依赖性GTPase活性。此外，针对重组Eo-eRF3的多克隆抗体被用于通过免疫荧光和免疫电子显微镜确定Eo-eRF3的定位，结果表明Eo-eRF3不仅分布在大核周围，还分布在八肋游仆虫细胞皮层的基体中。结果表明，Eo-eRF3除了在翻译终止中发挥作用外，可能还参与细胞骨架的组织。
