title: Pbp2x localizes separately from Pbp2b and other peptidoglycan synthesis proteins during later stages of cell division of Streptococcus pneumoniae D39.
abstract: The relative localization patterns of class B penicillin-binding proteins Pbp2x and Pbp2b were used as positional indicators of septal and peripheral (side-wall-like) peptidoglycan (PG) synthesis, respectively, in the mid-cell regions of Streptococcus pneumoniae cells at different stages of division. We confirm that Pbp2x and Pbp2b are essential in the strain D39 genetic background, which differs from that of laboratory strains. We show that Pbp2b, like Pbp2x and class A Pbp1a, follows a different localization pattern than FtsZ and remains at division septa after FtsZ reappears at the equators of daughter cells. Pulse-experiments with fluorescent D-amino acids (FDAAs) were performed in wild-type cells and in cells in which Pbp2x activity was preferentially inhibited by methicillin or Pbp2x amount was depleted. These experiments show that Pbp2x activity separates from that of other PBPs to the centres of constricting septa in mid-to-late divisional cells resolved by high-resolution 3D-SIM microscopy. Dual-protein and protein-fluorescent vancomycin 2D and 3D-SIM immunofluorescence microscopy (IFM) of cells at different division stages corroborate that Pbp2x separates to the centres of septa surrounded by an adjacent constricting ring containing Pbp2b, Pbp1a and regulators, StkP and MreC. The separate localization of Pbp2x suggests distinctive roles in completing septal PG synthesis and remodelling.
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, IN 47405.
authors.forename: Ho-Ching T
authors.initials: HT
authors.lastname: Tsui
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, IN 47405.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Boersma
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, IN 47405.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Vella
authors.affiliation: Department of Molecular and Cellular Biochemistry, Indiana University Bloomington, Bloomington, IN 47405.
authors.forename: Ozden
authors.initials: O
authors.lastname: Kocaoglu
authors.affiliation: Department of Molecular and Cellular Biochemistry, Indiana University Bloomington, Bloomington, IN 47405.
authors.forename: Erkin
authors.initials: E
authors.lastname: Kuru
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, IN 47405.
authors.forename: Julia K
authors.initials: JK
authors.lastname: Peceny
authors.affiliation: Department of Molecular and Cellular Biochemistry, Indiana University Bloomington, Bloomington, IN 47405.|Department of Chemistry, Indiana University Bloomington, Bloomington, IN 47405.
authors.forename: Erin E
authors.initials: EE
authors.lastname: Carlson
authors.affiliation: Department of Molecular and Cellular Biochemistry, Indiana University Bloomington, Bloomington, IN 47405.|Department of Chemistry, Indiana University Bloomington, Bloomington, IN 47405.
authors.forename: Michael S
authors.initials: MS
authors.lastname: VanNieuwenhze
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, IN 47405.|Department of Molecular and Cellular Biochemistry, Indiana University Bloomington, Bloomington, IN 47405.
authors.forename: Yves V
authors.initials: YV
authors.lastname: Brun
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, IN 47405.
authors.forename: Sidney L
authors.initials: SL
authors.lastname: Shaw
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, IN 47405.|Department of Molecular and Cellular Biochemistry, Indiana University Bloomington, Bloomington, IN 47405.
authors.forename: Malcolm E
authors.initials: ME
authors.lastname: Winkler
mesh_terms: D002455:Cell Division; D046915:Penicillin-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D010457:Peptidoglycan / Q000096:biosynthesis*; D021381:Protein Transport; D013296:Streptococcus pneumoniae / Q000166:cytology* / Q000235:genetics / Q000378:metabolism
chemical_list: D046915:Penicillin-Binding Proteins; D010457:Peptidoglycan; C072223:PBP 2x protein, Streptococcus
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