Loss of DNA-membrane interactions and cessation of DNA synthesis in myeloperoxidase-treated Escherichia coli.
Neutrophils and monocytes employ a diverse array of antimicrobial effector systems to support their host defense functions. The mechanisms of action of most of these systems are incompletely understood. The present report indicates that microbicidal activity by a neutrophil-derived antimicrobial system, consisting of myeloperoxidase, enzymatically generated hydrogen peroxide, and chloride ion, is accompanied by prompt cessation of DNA synthesis in Escherichia coli, as determined by markedly reduced incorporation of [3H]thymidine into trichloracetic acid-precipitable material. Simultaneously, the myeloperoxidase system mediates a decline in the ability of E. coli membranes to bind hemimethylated DNA sequences containing the E. coli chromosomal origin of replication (oriC). Binding of oriC to the E. coli membrane is an essential element of orderly chromosomal DNA replication. Comparable early changes in DNA synthesis and DNA-membrane interactions were not observed with alternative oxidant or antibiotic-mediated microbicidal systems. It is proposed that oxidants generated by the myeloperoxidase system modify the E. coli membrane in such a fashion that oriC binding is markedly impaired. As a consequence chromosomal DNA replication is impaired and organisms can no longer replicate.
Rosen
H
H
Department of Medicine, University of Washington, Seattle.
Orman
J
J
Rakita
R M
RM
Michel
B R
BR
VanDevanter
D R
DR
D002462:Cell Membrane / Q000378:metabolism; D002876:Chromosomes, Bacterial; D004261:DNA Replication / Q000187:drug effects*; D004269:DNA, Bacterial / Q000096:biosynthesis / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000254:growth & development*; D006801:Humans; D007700:Kinetics; D008745:Methylation; D009195:Peroxidase / Q000494:pharmacology*; D013936:Thymidine / Q000378:metabolism; D014316:Tritium
D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D014316:Tritium; D009195:Peroxidase; D013936:Thymidine