Cell wall glycans mediate recognition of the dairy bacterium Streptococcus thermophilus by bacteriophages

Paula Szymczak, Sérgio Raposo Filipe, Gonçalo Covas, Finn Kvist Vogensen, Ana Rute Neves, Thomas Janzen

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Receptors on the cell surfaces of bacterial hosts are essential during the infection cycle of bacteriophages. To date, the phage receptors of the industrial relevant dairy starter bacterium Streptococcus thermophilus remain elusive. Thus, we set out to identify cell surface structures that are involved in host recognition by dairy streptococcal phages. Five industrial S. thermophilus strains sensitive to different phages (pac type, cos type, and the new type 987), were selected to generate spontaneous bacteriophage-insensitive mutants (BIMs). Of these, approximately 50% were deselected as clustered regularly interspaced short palindromic repeat (CRISPR) mutants, while the other pool was further characterized to identify receptor mutants. On the basis of genome sequencing data, phage resistance in putative receptor mutants was attributed to nucleotide changes in genes encoding glycan biosynthetic pathways. Superresolution structured illumination microscopy was used to visualize the interactions between S. thermophilus and its phages. The phages were either regularly distributed along the cells or located at division sites of the cells. The cell wall structures mediating the latter type of phage adherence were further analyzed via phenotypic and biochemical assays. Altogether, our data suggested that phage adsorption to S. thermophilus is mediated by glycans associated with the bacterial cell surface. Specifically, the pac-type phage CHPC951 adsorbed to polysaccharides anchored to peptidoglycan, while the 987-type phage CHPC926 recognized exocellular polysaccharides associated with the cell surface.

Original languageEnglish
Article numbere01847-18
JournalApplied and Environmental Microbiology
Volume84
Issue number23
DOIs
Publication statusPublished - 1 Dec 2018

Fingerprint

Streptococcus Phages
Streptococcus thermophilus
bacteriophage
bacteriophages
Bacteriophages
Cell Wall
Polysaccharides
dairies
polysaccharides
cell walls
Bacteria
bacterium
bacteria
polysaccharide
mutants
microscopy
genome
cells
assay
receptors

Keywords

  • Adsorption
  • Bacteriophages
  • Cell wall
  • Glycans
  • Polysaccharides
  • Receptors
  • Streptococcus thermophilus

Cite this

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title = "Cell wall glycans mediate recognition of the dairy bacterium Streptococcus thermophilus by bacteriophages",
abstract = "Receptors on the cell surfaces of bacterial hosts are essential during the infection cycle of bacteriophages. To date, the phage receptors of the industrial relevant dairy starter bacterium Streptococcus thermophilus remain elusive. Thus, we set out to identify cell surface structures that are involved in host recognition by dairy streptococcal phages. Five industrial S. thermophilus strains sensitive to different phages (pac type, cos type, and the new type 987), were selected to generate spontaneous bacteriophage-insensitive mutants (BIMs). Of these, approximately 50{\%} were deselected as clustered regularly interspaced short palindromic repeat (CRISPR) mutants, while the other pool was further characterized to identify receptor mutants. On the basis of genome sequencing data, phage resistance in putative receptor mutants was attributed to nucleotide changes in genes encoding glycan biosynthetic pathways. Superresolution structured illumination microscopy was used to visualize the interactions between S. thermophilus and its phages. The phages were either regularly distributed along the cells or located at division sites of the cells. The cell wall structures mediating the latter type of phage adherence were further analyzed via phenotypic and biochemical assays. Altogether, our data suggested that phage adsorption to S. thermophilus is mediated by glycans associated with the bacterial cell surface. Specifically, the pac-type phage CHPC951 adsorbed to polysaccharides anchored to peptidoglycan, while the 987-type phage CHPC926 recognized exocellular polysaccharides associated with the cell surface.",
keywords = "Adsorption, Bacteriophages, Cell wall, Glycans, Polysaccharides, Receptors, Streptococcus thermophilus",
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Cell wall glycans mediate recognition of the dairy bacterium Streptococcus thermophilus by bacteriophages. / Szymczak, Paula; Filipe, Sérgio Raposo; Covas, Gonçalo; Vogensen, Finn Kvist; Neves, Ana Rute; Janzen, Thomas.

In: Applied and Environmental Microbiology, Vol. 84, No. 23, e01847-18, 01.12.2018.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Cell wall glycans mediate recognition of the dairy bacterium Streptococcus thermophilus by bacteriophages

AU - Szymczak, Paula

AU - Filipe, Sérgio Raposo

AU - Covas, Gonçalo

AU - Vogensen, Finn Kvist

AU - Neves, Ana Rute

AU - Janzen, Thomas

N1 - This work is part of an Industrial PhD project funded by Innovation Fund Denmark and Chr. Hansen A/S, grant no. 4135-00104B. Sem PDF conforme despacho.

PY - 2018/12/1

Y1 - 2018/12/1

N2 - Receptors on the cell surfaces of bacterial hosts are essential during the infection cycle of bacteriophages. To date, the phage receptors of the industrial relevant dairy starter bacterium Streptococcus thermophilus remain elusive. Thus, we set out to identify cell surface structures that are involved in host recognition by dairy streptococcal phages. Five industrial S. thermophilus strains sensitive to different phages (pac type, cos type, and the new type 987), were selected to generate spontaneous bacteriophage-insensitive mutants (BIMs). Of these, approximately 50% were deselected as clustered regularly interspaced short palindromic repeat (CRISPR) mutants, while the other pool was further characterized to identify receptor mutants. On the basis of genome sequencing data, phage resistance in putative receptor mutants was attributed to nucleotide changes in genes encoding glycan biosynthetic pathways. Superresolution structured illumination microscopy was used to visualize the interactions between S. thermophilus and its phages. The phages were either regularly distributed along the cells or located at division sites of the cells. The cell wall structures mediating the latter type of phage adherence were further analyzed via phenotypic and biochemical assays. Altogether, our data suggested that phage adsorption to S. thermophilus is mediated by glycans associated with the bacterial cell surface. Specifically, the pac-type phage CHPC951 adsorbed to polysaccharides anchored to peptidoglycan, while the 987-type phage CHPC926 recognized exocellular polysaccharides associated with the cell surface.

AB - Receptors on the cell surfaces of bacterial hosts are essential during the infection cycle of bacteriophages. To date, the phage receptors of the industrial relevant dairy starter bacterium Streptococcus thermophilus remain elusive. Thus, we set out to identify cell surface structures that are involved in host recognition by dairy streptococcal phages. Five industrial S. thermophilus strains sensitive to different phages (pac type, cos type, and the new type 987), were selected to generate spontaneous bacteriophage-insensitive mutants (BIMs). Of these, approximately 50% were deselected as clustered regularly interspaced short palindromic repeat (CRISPR) mutants, while the other pool was further characterized to identify receptor mutants. On the basis of genome sequencing data, phage resistance in putative receptor mutants was attributed to nucleotide changes in genes encoding glycan biosynthetic pathways. Superresolution structured illumination microscopy was used to visualize the interactions between S. thermophilus and its phages. The phages were either regularly distributed along the cells or located at division sites of the cells. The cell wall structures mediating the latter type of phage adherence were further analyzed via phenotypic and biochemical assays. Altogether, our data suggested that phage adsorption to S. thermophilus is mediated by glycans associated with the bacterial cell surface. Specifically, the pac-type phage CHPC951 adsorbed to polysaccharides anchored to peptidoglycan, while the 987-type phage CHPC926 recognized exocellular polysaccharides associated with the cell surface.

KW - Adsorption

KW - Bacteriophages

KW - Cell wall

KW - Glycans

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M3 - Article

VL - 84

JO - Applied and Environmental Microbiology

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