title: RatA (YfjG), an Escherichia coli toxin, inhibits 70S ribosome association to block translation initiation.
abstract: RatA (YfjG) is a toxin encoded by the ratA-ratB (yfjG-yfjF) operon on the Escherichia coli genome. Induction of RatA led to the inhibition of protein synthesis, while DNA and RNA synthesis was not affected. The stability of mRNAs was also unchanged as judged by in vivo primer extension experiments and by Northern blotting analysis. The ribosome profile of the cells overexpressing RatA showed that 70S ribosomes as well as polysomes significantly decreased with concomitant increase of 50S and 30S subunits. The addition of purified RatA to a cell-free system inhibited the formation of 70S ribosomes even in the presence of 6 mM Mg(2+) . RatA was specifically associated with 50S subunits, indicating that it binds to 50S subunits to block its association with 30S subunits leading to the inhibition of formation of 70S ribosomes. However, RatA did not cause dissociation of 70S ribosomes and its anti-association activity was blocked by paromomycin, an inhibitor for IF3, an essential initiation factor, having 21% sequence homology with RatA. Here we demonstrate that RatA is a new E. coli toxin, which effectively blocks the translation initiation step. We propose that this toxin of previously unknown function be renamed as RatA (Ribosome association toxin A).
authors.affiliation: Center for Advanced Biotechnology and Medicine, Department of Biochemistry, Robert Wood Johnson Medical School, 679 Hoes Lane, Piscataway, NJ 08854, USA.
authors.forename: Yonglong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Masayori
authors.initials: M
authors.lastname: Inouye
mesh_terms: D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D014176:Protein Biosynthesis; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism*
chemical_list: D001427:Bacterial Toxins; D029968:Escherichia coli Proteins; C558779:RatA protein, E coli