TY - JOUR
T1 - Function of bacteriophage G7C esterase tailspike in host cell adsorption
AU - Prokhorov, Nikolai S.
AU - Riccio, Cristian
AU - Zdorovenko, Evelina L.
AU - Shneider, Mikhail M.
AU - Browning, Christopher
AU - Knirel, Yuriy A.
AU - Leiman, Petr G.
AU - Letarov, Andrey V.
N1 - Funding Information:
We thank Prof. Jamila Horabin from the Florida State University for providing plasmid pDS3, Hannes Richter from the Genomics Technologies Facility at the Center for Integrative Genomics of the University of Lausanne for letting us use the Applied Biosystems 7900HT qPCR machine for the thermal stability experiment, Alexander S. Shashkov from the Zelinsky Institute for help with NMR spectroscopy. The work of the laboratory in the Winogradsky Institute was partially supported by Russian Science Foundation (RSF) grant #15?15-00134 (generation of the active site mutants, mapping of the gp63.1-gp66 interaction site, phage inactivation and adsorption inhibition assays, the bioinformatic work, the isolation and structure elucidation of the O-polysaccharide). The crystallographic experiments were performed on the X06SA beamline at the Swiss Light Source, Paul Scherrer Institut, Villigen, Switzerland. We thank Dr. Meitian Wang, Dr. Takashi Tomizaki and Dr. Vincent Olieric for their continuous support of the protein crystallography beamlines. The authors declare no conflict of interest.
PY - 2017/8
Y1 - 2017/8
N2 - Bacteriophages recognize and bind to their hosts with the help of receptor-binding proteins (RBPs) that emanate from the phage particle in the form of fibers or tailspikes. RBPs show a great variability in their shapes, sizes, and location on the particle. Some RBPs are known to depolymerize surface polysaccharides of the host while others show no enzymatic activity. Here we report that both RBPs of podovirus G7C – tailspikes gp63.1 and gp66 – are essential for infection of its natural host bacterium E. coli 4s that populates the equine intestinal tract. We characterize the structure and function of gp63.1 and show that unlike any previously described RPB, gp63.1 deacetylates surface polysaccharides of E. coli 4s leaving the backbone of the polysaccharide intact. We demonstrate that gp63.1 and gp66 form a stable complex, in which the N-terminal part of gp66 serves as an attachment site for gp63.1 and anchors the gp63.1-gp66 complex to the G7C tail. The esterase domain of gp63.1 as well as domains mediating the gp63.1-gp66 interaction is widespread among all three families of tailed bacteriophages.
AB - Bacteriophages recognize and bind to their hosts with the help of receptor-binding proteins (RBPs) that emanate from the phage particle in the form of fibers or tailspikes. RBPs show a great variability in their shapes, sizes, and location on the particle. Some RBPs are known to depolymerize surface polysaccharides of the host while others show no enzymatic activity. Here we report that both RBPs of podovirus G7C – tailspikes gp63.1 and gp66 – are essential for infection of its natural host bacterium E. coli 4s that populates the equine intestinal tract. We characterize the structure and function of gp63.1 and show that unlike any previously described RPB, gp63.1 deacetylates surface polysaccharides of E. coli 4s leaving the backbone of the polysaccharide intact. We demonstrate that gp63.1 and gp66 form a stable complex, in which the N-terminal part of gp66 serves as an attachment site for gp63.1 and anchors the gp63.1-gp66 complex to the G7C tail. The esterase domain of gp63.1 as well as domains mediating the gp63.1-gp66 interaction is widespread among all three families of tailed bacteriophages.
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U2 - 10.1111/mmi.13710
DO - 10.1111/mmi.13710
M3 - Article
C2 - 28513100
AN - SCOPUS:85021295736
VL - 105
SP - 385
EP - 398
JO - Molecular Microbiology
JF - Molecular Microbiology
SN - 0950-382X
IS - 3
ER -