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Ribosome rescue by Escherichia coli ArfA (YhdL) in the absence of trans-translation system.

Abstract：

:Although SsrA(tmRNA)-mediated trans-translation is thought to maintain the translation capacity of bacterial cells by rescuing ribosomes stalled on messenger RNA lacking an in-frame stop codon, single disruption of ssrA does not crucially hamper growth of Escherichia coli. Here, we identified YhdL (renamed ArfA for alternative ribosome-rescue factor) as a factor essential for the viability of E. coli in the absence of SsrA. The ssrA-arfA synthetic lethality was alleviated by SsrA(DD) , an SsrA variant that adds a proteolysis-refractory tag through trans-translation, indicating that ArfA-deficient cells require continued translation, rather than subsequent proteolysis of the truncated polypeptide. In accordance with this notion, depletion of SsrA in the ΔarfA background led to reduced translation of a model protein without affecting transcription, and puromycin, a codon-independent mimic of aminoacyl-tRNA, rescued the bacterial growth under such conditions. That ArfA takes over the role of SsrA was suggested by the observation that its overexpression enabled detection of the polypeptide encoded by a model non-stop mRNA, which was otherwise SsrA-tagged and degraded. In vitro, purified ArfA acted on a ribosome-nascent chain complex to resolve the peptidyl-tRNA. These results indicate that ArfA rescues the ribosome stalled at the 3' end of a non-stop mRNA without involving trans-translation.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

Chadani Y,Ono K,Ozawa S,Takahashi Y,Takai K,Nanamiya H,Tozawa Y,Kutsukake K,Abo T

doi

10.1111/j.1365-2958.2010.07375.x

subject

Has Abstract

pub_date

2010-11-01 00:00:00

pages

796-808

issue

4

eissn

0950-382X

issn

1365-2958

journal_volume

78

pub_type

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