The mechanism of plasmid curing in bacteria.
Bacterial plasmids have a major impact on metabolic function. Lactose fermentation of E. coli or hemolysin B transporter expressed by the plasmids that carry these respective genes could be readily obviated by heterocyclic compounds that readily bind to plasmid DNA. These compounds could also reverse the resistance to antibiotics of E. coli, Enterobacter, Proteus, Staphylococcus and Yersinia strains by eliminating plasmids. However, the frequency and extent of this effect was significantly less than might have been expected based on a complex interaction with plasmid DNA. The effects of heterocyclic compounds on the plasmids responsible for the virulence of Yersinia and A. tumefaciens, or on nodulation, nitrogen fixation of Rhizobia accounted for the elimination of 0.1 to 1.0 % of plasmids present in the populations studied. Bacterial plasmids can be eliminated from bacterial species grown as pure or mixed bacterial cultures in the presence of sub-inhibitory concentrations of non-mutagenic heterocyclic compounds. The antiplasmid action of the compounds depends on the chemical structure of amphiphillic compounds having a planar ring system with substitution in the L-molecular region. A symmetrical pi-electron conjugation at the highest occupied molecular orbitals favours the antiplasmid effect. The antiplasmid effect of heterocyclic compounds is expressed differentially in accordance with the structural form of the DNA to which they bind. In this manner "extrachromosomal" plasmid DNA that exists in a superhelical state binds more compound than its linear or open-circular form; and least to the chromosomal DNA of the bacterium, that carries the plasmid. It can also be noted that these compounds are not mutagenic and their antiplasmid effects correlate with the energy of HOMO-orbitals. Plasmid elimination is considered also to take place in ecosystems containing numerous bacterial species. This opens up a new perspective in rational drug design against bacterial plasmids. The inhibition of conjugational transfer of antibiotic resistance plasmid can be exploited to reduce the spread of antibiotic resistance plasmid in the ecosystem. Inhibition of plasmid replication at various stages, as shown in the "rolling circle" model (replication, partition, conjugal transfer) may also be the theoretical basis for the elimination of bacterial virulence in the case of plasmid mediated pathogenicity and antibiotic resistance. The large number of compounds tested for antiplasmid effects provides opportunities for QSAR studies in order to find a correlation between the antiplasmid effect and the supramolecular chemistry of these plasmid curing compounds. Plasmid elimination in vitro provides a method of isolating plasmid free bacteria for biotechnology without any risk of inducing mutations.
Spengler
Gabriella
G
Institute of Medical Microbiology and Immunobiology, Albert Szent-Györgyi Medical Centre, University of Szeged, Szeged, Hungary. spenglerg@freemail.hu
Molnár
Annamária
A
Schelz
Zsuzsanna
Z
Amaral
Leonard
L
Sharples
Derek
D
Molnár
Joseph
J
D001419:Bacteria / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D004261:DNA Replication / Q000187:drug effects / Q000235:genetics; D015195:Drug Design; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D006571:Heterocyclic Compounds / Q000737:chemistry / Q000494:pharmacology; D007364:Intercalating Agents / Q000737:chemistry / Q000494:pharmacology*; D015394:Molecular Structure; D010957:Plasmids / Q000235:genetics*; D021281:Quantitative Structure-Activity Relationship
D006571:Heterocyclic Compounds; D007364:Intercalating Agents