@article{BroekerKieleCasjensetal.2018,
  author    = {Broeker, Nina K. and Kiele, Franziska and Casjens, Sherwood R. and Gilcrease, Eddie B. and Thalhammer, Anja and Koetz, Joachim},
  title     = {In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620},
  series = {Viruses},
  volume    = {10},
  journal   = {Viruses},
  number    = {6},
  publisher = {Molecular Diversity Preservation International (MDPI)},
  address   = {Basel},
  issn      = {1999-4915},
  doi       = {10.3390/v10060289},
  pages     = {1 -- 15},
  year      = {2018},
  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.},
  language  = {en}
}
@article{StephanBroekerSaragliadisetal.2020,
  author    = {Stephan, Mareike Sophia and Br{\"o}ker, Nina K. and Saragliadis, Athanasios and Roos, Norbert and Linke, Dirk and Barbirz, Stefanie},
  title     = {In vitro analysis of O-antigen-specific bacteriophage P22 inactivation by Salmonella outer membrane vesicles},
  series = {Frontiers in microbiology},
  volume    = {11},
  journal   = {Frontiers in microbiology},
  publisher = {Frontiers Media},
  address   = {Lausanne},
  issn      = {1664-302X},
  doi       = {10.3389/fmicb.2020.510638},
  pages     = {12},
  year      = {2020},
  abstract  = {Bacteriophages use a large number of different bacterial cell envelope structures as receptors for surface attachment. As a consequence, bacterial surfaces represent a major control point for the defense against phage attack. One strategy for phage population control is the production of outer membrane vesicles (OMVs). In Gram-negative host bacteria, O-antigen-specific bacteriophages address lipopolysaccharide (LPS) to initiate infection, thus relying on an essential outer membrane glycan building block as receptor that is constantly present also in OMVs. In this work, we have analyzed interactions ofSalmonella(S.) bacteriophage P22 with OMVs. For this, we isolated OMVs that were formed in large amounts during mechanical cell lysis of the P22 S. Typhimurium host.In vitro, these OMVs could efficiently reduce the number of infective phage particles. Fluorescence spectroscopy showed that upon interaction with OMVs, bacteriophage P22 released its DNA into the vesicle lumen. However, only about one third of the phage P22 particles actively ejected their genome. For the larger part, no genome release was observed, albeit the majority of phages in the system had lost infectivity towards their host. With OMVs, P22 ejected its DNA more rapidly and could release more DNA against elevated osmotic pressures compared to DNA release triggered with protein-free LPS aggregates. This emphasizes that OMV composition is a key feature for the regulation of infective bacteriophage particles in the system.},
  language  = {en}
}