title: [Identification of a novel termination release factor eRF3b expressing the eRF3 activity in vitro and in vivo].
abstract: Termination of translation in eukaryotes is governed by the ribosome, a termination codon in the mRNA, and two polypeptide chain release factors (eRF1 and eRF3). We have identified a human protein of 628 amino acids, named eRF3b, which is highly homologous to the known human eRF3 henceforth named eRF3a. At the nucleotide and at the amino acid levels the human eRF3a and eRF3b are about 87% identical. The differences in amino acid sequence are concentrated near the amino terminus. The most important difference in the nucleotide sequence is that eRF3b lacks a GGC repeat close to the initiation codon in eRF3a. We have cloned the cDNA encoding the human eRF3b, purified the eRF3b expressed in Escherichia coli, and found that the protein is active in vitro as a potent stimulator of the release factor activity of human eRFl. Like eRF3a, eRF3b exhibits GTPase activity, which is ribosome- and eRFl-dependent. In vivo assays (based on suppression of readthrough induced by three species of suppressor tRNAs: amber, ochre, and opal) show that the human eRF3b is able to enhance the release factor activity of endogenous and overexpressed eRFl with all three stop codons.
authors.affiliation: Department of Molecular and Structural Biology, Aarhus University, 8000, Aarhus C, Denmark.
authors.forename: C G
authors.initials: CG
authors.lastname: Jakobsen
authors.forename: T M
authors.initials: TM
authors.lastname: Segaard
authors.forename: O
authors.initials: O
authors.lastname: Jean-Jean
authors.forename: L
authors.initials: L
authors.lastname: Frolova
authors.forename: J
authors.initials: J
authors.lastname: Justesen
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D004926:Escherichia coli; D020558:GTP Phosphohydrolases / Q000032:analysis / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010443:Peptide Chain Termination, Translational; D010454:Peptide Termination Factors / Q000032:analysis / Q000235:genetics*; D016415:Sequence Alignment
chemical_list: D018076:DNA, Complementary; D010454:Peptide Termination Factors; C087971:peptide-chain-release factor 3; D020558:GTP Phosphohydrolases
vernacular_title: Identifikatsiia novogo faktora terminatsii transliatsii eRF3b, obladaiushchego in vitro i in vivo aktivnost'iu eRF3.