Penicillin-binding protein 2 inactivation in Escherichia coli results in cell division inhibition, which is relieved by FtsZ overexpression.
Aminoacyl-tRNA synthetase mutants of Escherichia coli are resistant to amdinocillin (mecillinam), a beta-lactam antibiotic which specifically binds penicillin-binding protein 2 (PBP2) and prevents cell wall elongation with concomitant cell death. The leuS(Ts) strain, in which leucyl-tRNA synthetase is temperature sensitive, was resistant to amdinocillin at 37 degrees C because of an increased guanosine 5'-diphosphate 3'-diphosphate (ppGpp) pool resulting from partial induction of the stringent response, but it was sensitive to amdinocillin at 25 degrees C. We constructed a leuS(Ts) delta (rodA-pbpA)::Kmr strain, in which the PBP2 structural gene is deleted. This strain grew as spherical cells at 37 degrees C but was not viable at 25 degrees C. After a shift from 37 to 25 degrees C, the ppGpp pool decreased and cell division was inhibited; the cells slowly carried out a single division, increased considerably in volume, and gradually lost viability. The cell division inhibition was reversible when the ppGpp pool increased at high temperature, but reversion required de novo protein synthesis, possibly of septation proteins. The multicopy plasmid pZAQ, overproducing the septation proteins FtsZ, FtsA, and FtsQ, conferred amdinocillin resistance on a wild-type strain and suppressed the cell division inhibition in the leuS(Ts) delta (rodA-pbpA)::Kmr strain at 25 degrees C. The plasmid pAQ, in which the ftsZ gene is inactivated, did not confer amdinocillin resistance. These results lead us to hypothesize that the nucleotide ppGpp activates ftsZ expression and thus couples cell division to protein synthesis.
Vinella
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Institut Jacques Monod, Centre National de la Recherche Scientifique, Université Paris 7, France.
Joseleau-Petit
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Thévenet
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Bouloc
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D'Ari
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R
D000560:Amdinocillin; D001426:Bacterial Proteins / Q000502:physiology*; D002352:Carrier Proteins; D002455:Cell Division; D003598:Cytoskeletal Proteins; D004926:Escherichia coli / Q000166:cytology* / Q000187:drug effects; D019204:GTP-Binding Proteins / Q000502:physiology; D006159:Guanosine Tetraphosphate / Q000378:metabolism; D006602:Hexosyltransferases / Q000502:physiology*; D007935:Leucine-tRNA Ligase / Q000235:genetics; D009097:Multienzyme Complexes / Q000502:physiology*; D002267:Muramoylpentapeptide Carboxypeptidase; D010403:Penicillin Resistance; D046915:Penicillin-Binding Proteins; D010458:Peptidyl Transferases / Q000502:physiology*
D001426:Bacterial Proteins; D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; C071525:FtsZ protein, Bacteria; D009097:Multienzyme Complexes; D046915:Penicillin-Binding Proteins; D006159:Guanosine Tetraphosphate; D010458:Peptidyl Transferases; D006602:Hexosyltransferases; D002267:Muramoylpentapeptide Carboxypeptidase; D019204:GTP-Binding Proteins; D007935:Leucine-tRNA Ligase; D000560:Amdinocillin