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Viruses 2018, 10(6), 289; https://doi.org/10.3390/v10060289

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

1
Physikalische Biochemie, Universität Potsdam, Karl-Liebknecht-Str. 24-25, 14476 Golm, Germany
2
Division of Microbiology and Immunology, Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT 84112, USA
3
Kolloidchemie, Universität Potsdam, Karl-Liebknecht-Str.24-25, Haus 25, 14476 Golm, Germany
*
Author to whom correspondence should be addressed.
Received: 30 April 2018 / Revised: 19 May 2018 / Accepted: 21 May 2018 / Published: 29 May 2018
(This article belongs to the Special Issue Phage-Host Interactions)
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Abstract

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. View Full-Text
Keywords: O-antigen specific phage; podovirus; HK620; lipopolysaccharide; in vitro particle opening; tailspike protein O-antigen specific phage; podovirus; HK620; lipopolysaccharide; in vitro particle opening; tailspike protein
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Broeker, N.K.; Kiele, F.; Casjens, S.R.; Gilcrease, E.B.; Thalhammer, A.; Koetz, J.; Barbirz, S. In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620. Viruses 2018, 10, 289.

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