title: Expression, characterization and immunolocalization of translation termination factor eRF3 in the ciliate Euplotes octocarinatus.
abstract: 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.
authors.affiliation: Institute of Biotechnology, Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi Univesity, Taiyuan, PR China.
authors.forename: Bao-Feng
authors.initials: BF
authors.lastname: Chai
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Ai-Hua
authors.initials: AH
authors.lastname: Liang
mesh_terms: D000818:Animals; D018389:Codon, Nonsense; D004926:Escherichia coli / Q000378:metabolism; D016812:Euplotes / Q000378:metabolism* / Q000648:ultrastructure; D007150:Immunohistochemistry; D046529:Microscopy, Electron, Transmission; D009154:Mutation; D010454:Peptide Termination Factors / Q000235:genetics / Q000378:metabolism*; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D018389:Codon, Nonsense; D010454:Peptide Termination Factors; D011994:Recombinant Proteins; C087971:peptide-chain-release factor 3