Toxin Kid uncouples DNA replication and cell division to enforce retention of plasmid R1 in Escherichia coli cells.
Worldwide dissemination of antibiotic resistance in bacteria is facilitated by plasmids that encode postsegregational killing (PSK) systems. These produce a stable toxin (T) and a labile antitoxin (A) conditioning cell survival to plasmid maintenance, because only this ensures neutralization of toxicity. Shortage of antibiotic alternatives and the link of TA pairs to PSK have stimulated the opinion that premature toxin activation could be used to kill these recalcitrant organisms in the clinic. However, validation of TA pairs as therapeutic targets requires unambiguous understanding of their mode of action, consequences for cell viability, and function in plasmids. Conflicting with widespread notions concerning these issues, we had proposed that the TA pair kis-kid (killing suppressor-killing determinant) might function as a plasmid rescue system and not as a PSK system, but this remained to be validated. Here, we aimed to clarify unsettled mechanistic aspects of Kid activation, and of the effects of this for kis-kid-bearing plasmids and their host cells. We confirm that activation of Kid occurs in cells that are about to lose the toxin-encoding plasmid, and we show that this provokes highly selective restriction of protein outputs that inhibits cell division temporarily, avoiding plasmid loss, and stimulates DNA replication, promoting plasmid rescue. Kis and Kid are conserved in plasmids encoding multiple antibiotic resistance genes, including extended spectrum β-lactamases, for which therapeutic options are scarce, and our findings advise against the activation of this TA pair to fight pathogens carrying these extrachromosomal DNAs.
Medical Research Council Cancer Cell Unit, Hutchison/MRC Research Centre, Cambridge CB2 0XZ, United Kingdom.
Belén
B
Pimentel
Radhika
R
Nair
Camino
C
Bermejo-Rodríguez
Mark A
MA
Preston
Chukwuma A
CA
Agu
Xindan
X
Wang
Juan A
JA
Bernal
David J
DJ
Sherratt
Guillermo
G
de la Cueva-Méndez
D001483:Base Sequence; D015153:Blotting, Western; D002455:Cell Division / Q000502:physiology*; D004261:DNA Replication / Q000502:physiology*; D024881:Drug Resistance, Bacterial / Q000235:genetics / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000235:genetics; D011815:R Factors / Q000378:metabolism / Q000502:physiology*; D017422:Sequence Analysis, DNA
D029968:Escherichia coli Proteins; D009841:Oligonucleotides; C473072:kid toxin protein, E coli plasmid R1
PemK; RNase; mRNA interferase; parD; plasmid stability