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Flagellar structures from the bacterium Caulobacter crescentus and implications for phage ϕCbK predation of multi-flagellin bacteria

Eric J. Montemayor, Nicoleta T. Ploscariu, Juan C. Sanchez, Daniel Parrell, Rebecca S. Dillard, Conrad W. Shebelut, Zunlong Ke, Ricardo C. Guerrero-Ferreira, View ORCID ProfileElizabeth R. Wright
doi: https://doi.org/10.1101/2020.07.27.223248
Eric J. Montemayor
aDepartment of Biochemistry, University of Wisconsin, Madison, WI 53706
bCryo-Electron Microscopy Research Center, Department of Biochemistry, University of Wisconsin, Madison, WI 53706
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Nicoleta T. Ploscariu
aDepartment of Biochemistry, University of Wisconsin, Madison, WI 53706
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Juan C. Sanchez
aDepartment of Biochemistry, University of Wisconsin, Madison, WI 53706
cBiophysics Graduate Program, University of Wisconsin, Madison, WI 53706
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Daniel Parrell
aDepartment of Biochemistry, University of Wisconsin, Madison, WI 53706
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Rebecca S. Dillard
dDivision of Infectious Diseases, Department of Pediatrics, Emory University School of Medicine, Children’s Healthcare of Atlanta, Atlanta, GA, 30322
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Conrad W. Shebelut
eDepartment of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA, 30322
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Zunlong Ke
dDivision of Infectious Diseases, Department of Pediatrics, Emory University School of Medicine, Children’s Healthcare of Atlanta, Atlanta, GA, 30322
fSchool of Biological Sciences, Georgia Institute of Technology, Atlanta, GA, 30332
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Ricardo C. Guerrero-Ferreira
dDivision of Infectious Diseases, Department of Pediatrics, Emory University School of Medicine, Children’s Healthcare of Atlanta, Atlanta, GA, 30322
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Elizabeth R. Wright
aDepartment of Biochemistry, University of Wisconsin, Madison, WI 53706
bCryo-Electron Microscopy Research Center, Department of Biochemistry, University of Wisconsin, Madison, WI 53706
cBiophysics Graduate Program, University of Wisconsin, Madison, WI 53706
dDivision of Infectious Diseases, Department of Pediatrics, Emory University School of Medicine, Children’s Healthcare of Atlanta, Atlanta, GA, 30322
gMorgridge Institute for Research, Madison, WI, 53715
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  • ORCID record for Elizabeth R. Wright
  • For correspondence: [email protected]
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ABSTRACT

Caulobacter crescentus is a gram-negative alpha-proteobacterium that commonly lives in oligotrophic fresh and salt-water environments. C. crescentus is a host to many bacteriophages, including ϕCbK and ϕCbK-like bacteriophages, which first adsorb to cells by interaction with the bacterial flagellum. It is commonly thought that the six paralogs of the flagellin gene present in C. crescentus are important for bacteriophage evasion. Here, we show that deletion of specific flagellins in C. crescentus can indeed attenuate ϕCbK adsorption efficiency, although no single deletion completely ablates ϕCbK adsorption. Thus, bacteriophage ϕCbK likely recognizes a common motif amongst the six known flagellins in C. crescentus with varying degrees of efficiency. Interestingly, we observe that most deletion strains still generate flagellar filaments, with the exception of a strain that contains only the most divergent flagellin, FljJ, or a strain that contains only FljN and FljO. To visualize the surface residues that are likely recognized by ϕCbK, we determined two high-resolution structures of the FljK filament, with and without an amino acid substitution that induces straightening of the filament. We observe post-translational modifications on conserved surface threonine residues of FljK that are likely O-linked glycans. The possibility of interplay between these modifications and ϕCbK adsorption is discussed. We also determined the structure of a filament composed of a heterogeneous mixture of FljK and FljL, the final resolution of which was limited to approximately 4.6 Å. Altogether, this work builds a platform for future investigation of how phage ϕCbK infects C. crescentus at the molecular level.

Competing Interest Statement

The authors have declared no competing interest.

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Flagellar structures from the bacterium Caulobacter crescentus and implications for phage ϕCbK predation of multi-flagellin bacteria
Eric J. Montemayor, Nicoleta T. Ploscariu, Juan C. Sanchez, Daniel Parrell, Rebecca S. Dillard, Conrad W. Shebelut, Zunlong Ke, Ricardo C. Guerrero-Ferreira, Elizabeth R. Wright
bioRxiv 2020.07.27.223248; doi: https://doi.org/10.1101/2020.07.27.223248
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Flagellar structures from the bacterium Caulobacter crescentus and implications for phage ϕCbK predation of multi-flagellin bacteria
Eric J. Montemayor, Nicoleta T. Ploscariu, Juan C. Sanchez, Daniel Parrell, Rebecca S. Dillard, Conrad W. Shebelut, Zunlong Ke, Ricardo C. Guerrero-Ferreira, Elizabeth R. Wright
bioRxiv 2020.07.27.223248; doi: https://doi.org/10.1101/2020.07.27.223248

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