Stop codon selection in eukaryotic translation termination: comparison of the discriminating potential between human and ciliate eRF1s.
During eukaryotic translation termination, eRF1 responds to three stop codons. However, in ciliates with variant genetic codes, only one or two codons function as a stop signal. To localize the region of ciliate eRF1 implicated in stop codon discrimination, we have constructed ciliate-human hybrid eRF1s by swapping regions of human eRF1 for the equivalent region of ciliate Euplotes eRF1. We have examined the formation of a cross-link between recombinant eRF1s and mRNA analogs containing the photoactivable 4-thiouridine (s(4)U) at the first position of stop and control sense codons. With human eRF1, this cross-link can be detected only when either stop or UGG codons are located in the ribosomal A site. Here we show that the cross-link of the Euplotes-human hybrid eRF1 is restricted to mRNAs containing UAG and UAA codons, and that the entire N-terminal domain of Euplotes eRF1 is involved in discriminating against UGA and UGG. On the basis of these results, we discuss the steps of the selection process that determine the accuracy of stop codon recognition in eukaryotes.
Chavatte
Laurent
L
Institut Jacques Monod, UMR 7592 CNRS-Universités Paris 7-Paris 6, France.
Kervestin
Stéphanie
S
Favre
Alain
A
Jean-Jean
Olivier
O
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001483:Base Sequence; D016798:Ciliophora; D018388:Codon, Terminator; D017931:DNA Primers; D006801:Humans; D006822:Hybrid Cells; D008969:Molecular Sequence Data; D010454:Peptide Termination Factors / Q000737:chemistry / Q000235:genetics*; D014176:Protein Biosynthesis; D017386:Sequence Homology, Amino Acid
D018388:Codon, Terminator; D017931:DNA Primers; C489002:ETF1 protein, human; D010454:Peptide Termination Factors