TY  - GEN
A1  - Broeker, Nina K.
A1  - Kiele, Franziska
A1  - Casjens, Sherwood R.
A1  - Gilcrease, Eddie B.
A1  - Thalhammer, Anja
A1  - Koetz, Joachim
T1  - In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620
T2  - Viruses
N2  - 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.
T3  - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 469 
KW  - O-antigen specific phage
KW  - podovirus
KW  - HK620
KW  - lipopolysaccharide
KW  - in vitro particle opening
KW  - tailspike protein
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-417493
ER  - 
TY  - JOUR
A1  - Broeker, Nina K.
A1  - Kiele, Franziska
A1  - Casjens, Sherwood R.
A1  - Gilcrease, Eddie B.
A1  - Thalhammer, Anja
A1  - Koetz, Joachim
T1  - In Vitro Studies of Lipopolysaccharide-Mediated DNA Release of Podovirus HK620
JF  - Viruses
N2  - 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.
KW  - O-antigen specific phage
KW  - podovirus
KW  - HK620
KW  - lipopolysaccharide
KW  - in vitro particle opening
KW  - tailspike protein
Y1  - 2018
U6  - https://doi.org/10.3390/v10060289
SN  - 1999-4915
VL  - 10
IS  - 6
SP  - 1
EP  - 15
PB  - Molecular Diversity Preservation International (MDPI)
CY  - Basel
ER  -