Expression, characterization and immunolocalization of translation termination factor eRF3 in the ciliate Euplotes octocarinatus

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.