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PMID:19124462

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Citation

Zhang, Y and Inouye, M (2009) The inhibitory mechanism of protein synthesis by YoeB, an Escherichia coli toxin. J. Biol. Chem. 284:6627-38

Abstract

YoeB is a toxin encoded by the yefM-yoeB antitoxin-toxin operon in the Escherichia coli genome. Here we show that YoeB, a highly potent protein synthesis inhibitor, specifically blocks translation initiation. In in vivo primer extension experiments using two different mRNAs, a major band was detected after YoeB induction at three bases downstream of the initiation codon at 2.5 min. An identical band was also detected in in vitro toeprinting experiments after the addition of YoeB to the reaction mixtures containing 70 S ribosomes and the same mRNAs, even in the absence of tRNA(f)(Met). Notably, this band was not detected in the presence of YoeB alone, indicating that YoeB by itself does not have endoribonuclease activity under the conditions used. The 70 S ribosomes increased upon YoeB induction, and YoeB was found to be specifically associated with 50 S subunits. Using tetracycline and hygromycin B, we demonstrated that YoeB binds to the 50 S ribosomal subunit in 70 S ribosomes and interacts with the A site leading to mRNA cleavage at this site. As a result, the 3'-end portion of the mRNA was released from ribosomes, and translation initiation was effectively inhibited. These results demonstrate that YoeB primarily inhibits translation initiation.

Links

PubMed PMC2652283 Online version:10.1074/jbc.M808779200

Keywords

Anti-Bacterial Agents/pharmacology; Bacterial Toxins/chemistry; Bacterial Toxins/metabolism; Cell-Free System/chemistry; Cell-Free System/metabolism; Codon, Initiator/chemistry; Codon, Initiator/metabolism; Escherichia coli/chemistry; Escherichia coli/metabolism; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/metabolism; Hygromycin B/pharmacology; Peptide Chain Initiation, Translational/drug effects; Peptide Chain Initiation, Translational/physiology; Protein Synthesis Inhibitors/chemistry; Protein Synthesis Inhibitors/metabolism; RNA, Bacterial/chemistry; RNA, Bacterial/metabolism; RNA, Transfer, Amino Acyl/chemistry; RNA, Transfer, Amino Acyl/metabolism; RNA, Transfer, Met/chemistry; RNA, Transfer, Met/metabolism; Ribosome Subunits, Large, Bacterial/chemistry; Ribosome Subunits, Large, Bacterial/metabolism; Tetracycline/pharmacology

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

ECOLI:YOEB

acts_upstream_of_or_within

GO:0045947: negative regulation of translational initiation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:YOEB

involved_in

GO:0045947: negative regulation of translational initiation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:YOEB

enables

GO:0043024: ribosomal small subunit binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

Notes

See also

References

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