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

Rasmus N. Klitgaard; Anders Løbner-Olesen

doi:10.2174/1570163815666180423115514

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

Rasmus N. Klitgaard, Anders Løbner-Olesen

Functional genomics

2 Citationer (Scopus)

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.

Originalsprog	Engelsk
Tidsskrift	Current Drug Discovery Technologies
Vol/bind	16
Udgave nummer	3
Sider (fra-til)	272-277
ISSN	1570-1638
DOI	https://doi.org/10.2174/1570163815666180423115514
Status	Udgivet - 2019

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10.2174/1570163815666180423115514

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title = "A Novel Fluorescence-Based Screen for Inhibitors of the Initiation of DNA Replication in Bacteria",

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.",

author = "Klitgaard, {Rasmus N.} and Anders L{\o}bner-Olesen",

note = "Copyright{\textcopyright} Bentham Science Publishers; For any queries, please email at [email protected].",

year = "2019",

doi = "10.2174/1570163815666180423115514",

language = "English",

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T1 - A Novel Fluorescence-Based Screen for Inhibitors of the Initiation of DNA Replication in Bacteria

AU - Klitgaard, Rasmus N.

AU - Løbner-Olesen, Anders

PY - 2019

Y1 - 2019

N2 - 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.

AB - 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.

U2 - 10.2174/1570163815666180423115514

DO - 10.2174/1570163815666180423115514

M3 - Journal article

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SN - 1570-1638

VL - 16

SP - 272

EP - 277

JO - Current Drug Discovery Technologies

JF - Current Drug Discovery Technologies

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ER -

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

Abstract

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