Nov 8, 2018

A new mechanism for ribosome rescue can recruit RF1 or RF2 to non-stop ribosomes

BioRxiv : the Preprint Server for Biology
Tyler D GoralskiKenneth C. Keiler

Abstract

Bacterial ribosomes frequently translate to the 3' end of an mRNA without terminating at an in-frame stop codon. In all bacteria studied to date, these non-stop ribosomes are rescued using trans-translation. In some species, genes required for trans-translation are essential, but other species can survive without trans-translation because they express an alternative ribosome rescue factor, ArfA or ArfB. Francisella tularensis cells lacking trans-translation are viable, but F. tularensis does not encode ArfA or ArfB. Transposon mutagenesis followed by deep sequencing (Tn-seq) identified a new alternative ribosome rescue factor, now named ArfT. arfT can be deleted in wild-type cells but not in cells that lack trans-translation activity. Over-expression of ArfT suppresses the slow growth phenotype in cells lacking trans-translation and counteracts growth arrest caused by trans-translation inhibitors, indicating that ArfT rescues non-stop ribosomes in vivo. Ribosome rescue assays in vitro show that ArfT promotes hydrolysis of peptidyl-tRNA on non-stop ribosomes in conjunction with F. tularensis release factors. Unlike ArfA, which requires RF2 for activity, ArfT can function with either RF1 or RF2. Overall, these results indicate th...Continue Reading

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Mentioned in this Paper

In Vivo
TRNA, peptidyl-
Genes
Trans-translation
Inhibitors
Codon, Terminator
Etiology
Francisella Tularensis Antibody Assay
ArfA protein, E coli
1-(9'-(o-carboxyphenyl)-6'-amino-2'-chloro-3'-xanthanone)-4,10-(diethyl)-7-(2-pyridylmethyl)-1,4,7,10-tetraazacyclododecane

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