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Inhibitors of Bacterial DNA Synthesis01:28

Inhibitors of Bacterial DNA Synthesis

Bacterial pathogens depend on precise and efficient DNA replication to sustain infection. Two type II topoisomerases—DNA gyrase and topoisomerase IV—are critical to this process, as they resolve DNA supercoiling and unlink chromosomes during replication. Fluoroquinolones, synthetic derivatives of quinolones, exploit this mechanism by stabilizing the transient DNA–enzyme cleavage complex, preventing strand religation, and causing lethal double-strand breaks. These antibiotics are selectively...
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Nanoparticles inhibit DNA replication by binding to DNA: modeling and experimental validation.

Kungang Li1, Xiaonan Zhao, Brian K Hammer

  • 1School of Civil and Environmental Engineering, Georgia Institute of Technology , Atlanta, Georgia 30332, United States.

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Predictive models and experimental methods can identify harmful nanoparticles (NPs). High NP-DNA binding affinity correlates with inhibited DNA replication, aiding in the design of safer nanomaterials.

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Area of Science:

  • Nanotechnology
  • Toxicology
  • Molecular Biology

Background:

  • Predictive models aid nanoparticle (NP) toxicological testing, but experimental validation is crucial for identifying harmful NPs.
  • Nanoparticle-DNA interactions are key to understanding NP toxicity, as they can interfere with DNA function.
  • Current methods for assessing NP-DNA interactions and subsequent toxicity are time-consuming and expensive.

Purpose of the Study:

  • To predict and experimentally determine the binding affinity of various nanoparticles (NPs) to DNA.
  • To investigate the effect of NP-DNA binding affinity on DNA replication in vitro.
  • To establish a methodology for efficient genotoxicological testing and safe NP design.

Main Methods:

  • Calculated NP-DNA interaction energy using the Derjaguin-Landau-Verwey-Overbeek (DLVO) model for 12 NP types.
  • Measured NP-DNA binding affinity experimentally using atomic force microscopy (AFM).
  • Assessed the impact of NPs on DNA replication using the polymerase chain reaction (PCR) technique.

Main Results:

  • Theoretical predictions of NP-DNA binding affinity closely matched experimental AFM observations.
  • NPs exhibiting high DNA binding affinity significantly inhibited in vitro DNA replication.
  • NPs with low DNA binding affinity demonstrated minimal or no impact on DNA replication.

Conclusions:

  • The combined theoretical and experimental approach accurately predicts NP-DNA binding affinity.
  • NP-DNA binding affinity is a strong indicator of potential genotoxicity, specifically affecting DNA replication.
  • This methodology offers a valuable tool for prioritizing NPs for toxicological testing and designing safer nanomaterials.