A Novel Family of Escherichia coli Toxin-Antitoxin Gene Pairs

ABSTRACT Bacterial toxin-antitoxin protein pairs (TA pairs) encode a toxin protein, which poisons cells by binding and inhibiting an essential enzyme, and an antitoxin protein, which binds the toxin and restores viability. We took an approach that did not rely on sequence homology to search for unidentified TA pairs in the genome of Escherichia coli K-12. Of 32 candidate genes tested, ectopic expression of 6 caused growth inhibition. In this report, we focus on the initial characterization of yeeV, ykfI, and ypjF, a novel family of toxin proteins. Coexpression of the gene upstream of each toxin restored the growth rate to that of the uninduced strain. Unexpectedly, we could not detect in vivo protein-protein interactions between the new toxin and antitoxin pairs. Instead, the antitoxins appeared to function by causing a large reduction in the level of cellular toxin protein.

[1]  D. Kennell,et al.  Translation and mRNA decay , 1978, Molecular and General Genetics MGG.

[2]  Jeroen A. A. Demmers,et al.  Interactions between Phage-Shock Proteins in Escherichia coli , 2003, Journal of bacteriology.

[3]  Måns Ehrenberg,et al.  The Bacterial Toxin RelE Displays Codon-Specific Cleavage of mRNAs in the Ribosomal A Site , 2003, Cell.

[4]  K. Gerdes,et al.  Rapid induction and reversal of a bacteriostatic condition by controlled expression of toxins and antitoxins , 2002, Molecular microbiology.

[5]  K. Gerdes,et al.  RelE, a global inhibitor of translation, is activated during nutritional stress , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[6]  Purification of the RelB and RelE Proteins ofEscherichia coli: RelE Binds to RelB and to Ribosomes , 2001, Journal of bacteriology.

[7]  V. L. Miller,et al.  The psp locus of Yersinia enterocolitica is required for virulence and for growth in vitro when the Ysc type III secretion system is produced , 2001, Molecular microbiology.

[8]  U. Zielenkiewicz,et al.  Mechanisms of plasmid stable maintenance with special focus on plasmid addiction systems. , 2001, Acta biochimica Polonica.

[9]  K. Lewis,et al.  Programmed Death in Bacteria , 2000, Microbiology and Molecular Biology Reviews.

[10]  K. Shaw,et al.  Evernimicin Binds Exclusively to the 50S Ribosomal Subunit and Inhibits Translation in Cell-Free Systems Derived from both Gram-Positive and Gram-Negative Bacteria , 2000, Antimicrobial Agents and Chemotherapy.

[11]  K. Gerdes.  Toxin-Antitoxin Modules May Regulate Synthesis of Macromolecules during Nutritional Stress , 2000, Journal of bacteriology.

[12]  G. Mittenhuber,et al.  Occurrence of mazEF-like antitoxin/toxin systems in bacteria. , 1999, Journal of molecular microbiology and biotechnology.

[13]  A. Darwin,et al.  Identification of Yersinia enterocolitica genes affecting survival in an animal host using signature‐tagged transposon mutagenesis , 1999, Molecular microbiology.

[14]  A. Bateman.  The SIS domain: a phosphosugar-binding domain. , 1999, Trends in biochemical sciences.

[15]  H. Engelberg-Kulka,et al.  Addiction modules and programmed cell death and antideath in bacterial cultures. , 1999, Annual review of microbiology.

[16]  F. Hayes.  A Family of Stability Determinants in Pathogenic Bacteria , 1998, Journal of bacteriology.

[17]  K. Gerdes,et al.  The Escherichia coli relBE genes belong to a new toxin–antitoxin gene family , 1998, Molecular microbiology.

[18]  N. W. Davis,et al.  The complete genome sequence of Escherichia coli K-12. , 1997, Science.

[19]  J. Dworkin,et al.  The Escherichia coli phage‐shock‐protein (psp) operon , 1997, Molecular microbiology.

[20]  H. Engelberg-Kulka,et al.  An Escherichia coli chromosomal "addiction module" regulated by guanosine [corrected] 3',5'-bispyrophosphate: a model for programmed bacterial cell death. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[21]  M. Valvano,et al.  Biosynthesis of Inner Core Lipopolysaccharide in Enteric Bacteria Identification and Characterization of a Conserved Phosphoheptose Isomerase (*) , 1996, The Journal of Biological Chemistry.

[22]  M. Kleerebezem,et al.  Involvement of stress protein PspA (phage shock protein A) of Escherichia coli in maintenance of the protonmotive force under stress conditions. , 1996, The EMBO journal.

[23]  D. Belin,et al.  Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter , 1995, Journal of bacteriology.

[24]  P. Model,et al.  Analysis of the proteins and cis-acting elements regulating the stress-induced phage shock protein operon. , 1995, Nucleic acids research.

[25]  M. Ruiz-Echevarría,et al.  Kid, a small protein of the parD stability system of plasmid R1, is an inhibitor of DNA replication acting at the initiation of DNA synthesis. , 1995, Journal of molecular biology.

[26]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[27]  H. Bergler,et al.  Inhibition of lipid biosynthesis induces the expression of the pspA gene. , 1994, Microbiology.

[28]  E. Ohtsubo,et al.  chpA and chpB, Escherichia coli chromosomal homologs of the pem locus responsible for stable maintenance of plasmid R100 , 1993, Journal of bacteriology.

[29]  M. Couturier,et al.  Cell killing by the F plasmid CcdB protein involves poisoning of DNA-topoisomerase II complexes. , 1992, Journal of molecular biology.

[30]  T. Miki,et al.  Control of segregation of chromosomal DNA by sex factor F in Escherichia coli. Mutants of DNA gyrase subunit A suppress letD (ccdB) product growth inhibition. , 1992, Journal of molecular biology.

[31]  P. Model,et al.  Stress-induced expression of the Escherichia coli phage shock protein operon is dependent on sigma 54 and modulated by positive and negative feedback mechanisms. , 1991, Genes & development.

[32]  P. Model,et al.  Characterization and sequence of the Escherichia coli stress-induced psp operon. , 1991, Journal of molecular biology.

[33]  S. Ho,et al.  Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction. , 2013, BioTechniques.

[34]  P. Model,et al.  Phage shock protein, a stress protein of Escherichia coli. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[35]  W. Fiers,et al.  Inefficient translation initiation causes premature transcription termination in the IacZ gene , 1986, Cell.

[36]  F. W. Bech,et al.  Sequence of the relB transcription unit from Escherichia coli and identification of the relB gene. , 1985, The EMBO journal.

[37]  B. Diderichsen,et al.  Genetics of the relB locus in Escherichia coli , 1977, Journal of bacteriology.

[38]  A. T.,et al.  On Stringent Response , 1972, Nature.

[39]  R. Lavallé.  [New mutants for regulation of RNA synthesis]. , 1965, Bulletin de la Societe de chimie biologique.