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Article

Exploiting a Phage-Bacterium Interaction System as a Molecular Switch to Decipher Macromolecular Interactions in the Living Cell

1
Department of Applied Biotechnology and Food Sciences, Budapest University of Technology and Economics, H-1111 Budapest, Hungary
2
Institute of Enzymology, RCNS, Hungarian Academy of Sciences, H-1111 Budapest, Hungary
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work as first authors.
Viruses 2018, 10(4), 168; https://doi.org/10.3390/v10040168
Received: 20 February 2018 / Revised: 22 March 2018 / Accepted: 30 March 2018 / Published: 1 April 2018
(This article belongs to the Special Issue Biotechnological Applications of Phage and Phage-Derived Proteins)
Pathogenicity islands of Staphylococcus aureus are under the strong control of helper phages, where regulation is communicated at the gene expression level via a family of specific repressor proteins. The repressor proteins are crucial to phage-host interactions and, based on their protein characteristics, may also be exploited as versatile molecular tools. The Stl repressor from this protein family has been recently investigated and although the binding site of Stl on DNA was recently discovered, there is a lack of knowledge on the specific protein segments involved in this interaction. Here, we develop a generally applicable system to reveal the mechanism of the interaction between Stl and its cognate DNA within the cellular environment. Our unbiased approach combines random mutagenesis with high-throughput analysis based on the lac operon to create a well-characterized gene expression system. Our results clearly indicate that, in addition to a previously implicated helix-turn-helix segment, other protein moieties also play decisive roles in the DNA binding capability of Stl. Structural model-based investigations provided a detailed understanding of Stl:DNA complex formation. The robustness and reliability of our novel test system were confirmed by several mutated Stl constructs, as well as by demonstrating the interaction between Stl and dUTPase from the Staphylococcal ϕ11 phage. Our system may be applied to high-throughput studies of protein:DNA and protein:protein interactions. View Full-Text
Keywords: gene expression regulation; molecular probe; macromolecular interactions; phage-host interaction gene expression regulation; molecular probe; macromolecular interactions; phage-host interaction
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MDPI and ACS Style

Surányi, É.V.; Hírmondó, R.; Nyíri, K.; Tarjányi, S.; Kőhegyi, B.; Tóth, J.; Vértessy, B.G. Exploiting a Phage-Bacterium Interaction System as a Molecular Switch to Decipher Macromolecular Interactions in the Living Cell. Viruses 2018, 10, 168. https://doi.org/10.3390/v10040168

AMA Style

Surányi ÉV, Hírmondó R, Nyíri K, Tarjányi S, Kőhegyi B, Tóth J, Vértessy BG. Exploiting a Phage-Bacterium Interaction System as a Molecular Switch to Decipher Macromolecular Interactions in the Living Cell. Viruses. 2018; 10(4):168. https://doi.org/10.3390/v10040168

Chicago/Turabian Style

Surányi, Éva V., Rita Hírmondó, Kinga Nyíri, Szilvia Tarjányi, Bianka Kőhegyi, Judit Tóth, and Beáta G. Vértessy 2018. "Exploiting a Phage-Bacterium Interaction System as a Molecular Switch to Decipher Macromolecular Interactions in the Living Cell" Viruses 10, no. 4: 168. https://doi.org/10.3390/v10040168

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