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In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620

Gram-negative bacteria protect themselves with an outermost layer containing lipopolysaccharide (LPS). O-antigen-specific bacteriophages use tailspike proteins (TSP) to recognize and cleave the O-polysaccharide part of LPS. However, O-antigen composition and structure can be highly variable dependin...

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Autores principales: Broeker, Nina K., Kiele, Franziska, Casjens, Sherwood R., Gilcrease, Eddie B., Thalhammer, Anja, Koetz, Joachim, Barbirz, Stefanie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6024685/
https://www.ncbi.nlm.nih.gov/pubmed/29843473
http://dx.doi.org/10.3390/v10060289
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author Broeker, Nina K.
Kiele, Franziska
Casjens, Sherwood R.
Gilcrease, Eddie B.
Thalhammer, Anja
Koetz, Joachim
Barbirz, Stefanie
author_facet Broeker, Nina K.
Kiele, Franziska
Casjens, Sherwood R.
Gilcrease, Eddie B.
Thalhammer, Anja
Koetz, Joachim
Barbirz, Stefanie
author_sort Broeker, Nina K.
collection PubMed
description Gram-negative bacteria protect themselves with an outermost layer containing lipopolysaccharide (LPS). O-antigen-specific bacteriophages use tailspike proteins (TSP) to recognize and cleave the O-polysaccharide part of LPS. However, O-antigen composition and structure can be highly variable depending on the environmental conditions. It is important to understand how these changes may influence the early steps of the bacteriophage infection cycle because they can be linked to changes in host range or the occurrence of phage resistance. In this work, we have analyzed how LPS preparations in vitro trigger particle opening and DNA ejection from the E. coli podovirus HK620. Fluorescence-based monitoring of DNA release showed that HK620 phage particles in vitro ejected their genome at velocities comparable to those found for other podoviruses. Moreover, we found that HK620 irreversibly adsorbed to the LPS receptor via its TSP at restrictive low temperatures, without opening the particle but could eject its DNA at permissive temperatures. DNA ejection was solely stimulated by LPS, however, the composition of the O-antigen dictated whether the LPS receptor could start the DNA release from E. coli phage HK620 in vitro. This finding can be significant when optimizing bacteriophage mixtures for therapy, where in natural environments O-antigen structures may rapidly change.
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spelling pubmed-60246852018-07-16 In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620 Broeker, Nina K. Kiele, Franziska Casjens, Sherwood R. Gilcrease, Eddie B. Thalhammer, Anja Koetz, Joachim Barbirz, Stefanie Viruses Article Gram-negative bacteria protect themselves with an outermost layer containing lipopolysaccharide (LPS). O-antigen-specific bacteriophages use tailspike proteins (TSP) to recognize and cleave the O-polysaccharide part of LPS. However, O-antigen composition and structure can be highly variable depending on the environmental conditions. It is important to understand how these changes may influence the early steps of the bacteriophage infection cycle because they can be linked to changes in host range or the occurrence of phage resistance. In this work, we have analyzed how LPS preparations in vitro trigger particle opening and DNA ejection from the E. coli podovirus HK620. Fluorescence-based monitoring of DNA release showed that HK620 phage particles in vitro ejected their genome at velocities comparable to those found for other podoviruses. Moreover, we found that HK620 irreversibly adsorbed to the LPS receptor via its TSP at restrictive low temperatures, without opening the particle but could eject its DNA at permissive temperatures. DNA ejection was solely stimulated by LPS, however, the composition of the O-antigen dictated whether the LPS receptor could start the DNA release from E. coli phage HK620 in vitro. This finding can be significant when optimizing bacteriophage mixtures for therapy, where in natural environments O-antigen structures may rapidly change. MDPI 2018-05-29 /pmc/articles/PMC6024685/ /pubmed/29843473 http://dx.doi.org/10.3390/v10060289 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Broeker, Nina K.
Kiele, Franziska
Casjens, Sherwood R.
Gilcrease, Eddie B.
Thalhammer, Anja
Koetz, Joachim
Barbirz, Stefanie
In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620
title In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620
title_full In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620
title_fullStr In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620
title_full_unstemmed In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620
title_short In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620
title_sort in vitro studies of lipopolysaccharide-mediated dna release of podovirus hk620
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6024685/
https://www.ncbi.nlm.nih.gov/pubmed/29843473
http://dx.doi.org/10.3390/v10060289
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