A Novel Fluorescence-Based Screen for Inhibitors of the Initiation of DNA Replication in Bacteria

Author(s): Rasmus N. Klitgaard, Anders Løbner-Olesen*.

Journal Name: Current Drug Discovery Technologies

Volume 16 , Issue 3 , 2019

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Graphical Abstract:


Abstract:

Background: One of many strategies to overcome antibiotic resistance is the discovery of compounds targeting cellular processes, which have not yet been exploited.

Materials and Methods: Using various genetic tools, we constructed a novel high throughput, cellbased, fluorescence screen for inhibitors of chromosome replication initiation in bacteria.

Results: The screen was validated by expression of an intra-cellular cyclic peptide interfering with the initiator protein DnaA and by over-expression of the negative initiation regulator SeqA. We also demonstrated that neither tetracycline nor ciprofloxacin triggers a false positive result. Finally, 400 extracts isolated mainly from filamentous actinomycetes were subjected to the screen.

Conclusion: We concluded that the presented screen is applicable for identifying putative inhibitors of DNA replication initiation in a high throughput setup.

Keywords: DNA replication initiation, inhibitors, DnaA, high throughput screen, fluorescence, microbial extracts.

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Article Details

VOLUME: 16
ISSUE: 3
Year: 2019
Page: [272 - 277]
Pages: 6
DOI: 10.2174/1570163815666180423115514
Price: $65

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