Please use this identifier to cite or link to this item: doi:10.22028/D291-43502
Title: Establishment of an In Bacterio Assay for the Assessment of Carbon Storage Regulator A (CsrA) Inhibitors
Author(s): Yingwen, Wu
Zoller, Ben G. E.
Kamal, Mohamed Ashraf Mostafa
Hotop, Sven-Kevin
Lehr, Claus-Michael
Brönstrup, Mark
Dersch, Petra
Empting, Martin
Language: English
Title: Chembiochem : a European journal of chemical biology
Volume: 24
Issue: 16
Publisher/Platform: Wiley
Year of Publication: 2023
DDC notations: 500 Science
Publikation type: Journal Article
Abstract: Polymicrobial infections involving various combinations of microorganisms, such as Escherichia, Pseudomonas, or Yersinia, can lead to acute and chronic diseases in for example the gastrointestinal and respiratory tracts. Our aim is to modulate microbial communities by targeting the posttranscriptional regulator system called carbon storage regulator A (CsrA) (or also repressor of secondary metabolites (RsmA)). In previous studies, we identified easily accessible CsrA binding scaffolds and macrocyclic CsrA binding peptides through biophysical screening and phage display technology. However, due to the lack of an appropriate in bacterio assay to evaluate the cellular effects of these inhibitor hits, the focus of the present study is to establish an in bacterio assay capable of probing and quantifying the impact on CsrA-regulated cellular mechanisms. We have successfully developed an assay based on a luciferase reporter gene assay, which in combination with a qPCR expression gene assay, allows for the monitoring of expression levels of different downstream targets of CsrA. The chaperone protein CesT was used as a suitable positive control for the assay, and in time-dependent experiments, we observed a CesT-mediated increase in bioluminescence over time. By this means, the cellular on-target effects of non-bactericidal/non-bacteriostatic virulence modulating compounds targeting CsrA/RsmA can be evaluated.
DOI of the first publication: 10.1002/cbic.202300369
URL of the first publication: https://chemistry-europe.onlinelibrary.wiley.com/doi/epdf/10.1002/cbic.202300369
Link to this record: urn:nbn:de:bsz:291--ds-435020
hdl:20.500.11880/38987
http://dx.doi.org/10.22028/D291-43502
ISSN: 1439-7633
1439-4227
Date of registration: 19-Nov-2024
Faculty: NT - Naturwissenschaftlich- Technische Fakultät
Department: NT - Biowissenschaften
NT - Pharmazie
Professorship: NT - Prof. Dr. Anna Hirsch
NT - Prof. Dr. Claus-Michael Lehr
NT - Keiner Professur zugeordnet
Collections:SciDok - Der Wissenschaftsserver der Universität des Saarlandes



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