Suppression of a deletion mutation in the gene encoding essential PBP2b reveals a new lytic transglycosylase involved in peripheral peptidoglycan synthesis in Streptococcus pneumoniae D39

Mol Microbiol. 2016 Jun;100(6):1039-65. doi: 10.1111/mmi.13366. Epub 2016 Apr 15.

Abstract

In ellipsoid-shaped ovococcus bacteria, such as the pathogen Streptococcus pneumoniae (pneumococcus), side-wall (peripheral) peptidoglycan (PG) synthesis emanates from midcells and is catalyzed by the essential class B penicillin-binding protein PBP2b transpeptidase (TP). We report that mutations that inactivate the pneumococcal YceG-domain protein, Spd_1346 (renamed MltG), remove the requirement for PBP2b. ΔmltG mutants in unencapsulated strains accumulate inactivation mutations of class A PBP1a, which possesses TP and transglycosylase (TG) activities. The 'synthetic viable' genetic relationship between Δpbp1a and ΔmltG mutations extends to essential ΔmreCD and ΔrodZ mutations that misregulate peripheral PG synthesis. Remarkably, the single MltG(Y488D) change suppresses the requirement for PBP2b, MreCD, RodZ and RodA. Structural modeling and comparisons, catalytic-site changes and an interspecies chimera indicate that pneumococcal MltG is the functional homologue of the recently reported MltG endo-lytic transglycosylase of Escherichia coli. Depletion of pneumococcal MltG or mltG(Y488D) increases sphericity of cells, and MltG localizes with peripheral PG synthesis proteins during division. Finally, growth of Δpbp1a ΔmltG or mltG(Y488D) mutants depends on induction of expression of the WalRK TCS regulon of PG hydrolases. These results fit a model in which MltG releases anchored PG glycan strands synthesized by PBP1a for crosslinking by a PBP2b:RodA complex in peripheral PG synthesis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Aminoacyltransferases / genetics*
  • Aminoacyltransferases / metabolism
  • Glycosyltransferases / metabolism
  • N-Acetylmuramoyl-L-alanine Amidase / metabolism
  • Penicillin-Binding Proteins / genetics*
  • Penicillin-Binding Proteins / metabolism
  • Peptidoglycan / biosynthesis*
  • Peptidoglycan / metabolism
  • Peptidyl Transferases / genetics
  • Peptidyl Transferases / metabolism
  • Sequence Deletion*
  • Streptococcus pneumoniae / enzymology
  • Streptococcus pneumoniae / genetics*
  • Streptococcus pneumoniae / metabolism

Substances

  • Penicillin-Binding Proteins
  • Peptidoglycan
  • Aminoacyltransferases
  • penicillin-binding protein 2b, Streptococcus
  • Peptidyl Transferases
  • Glycosyltransferases
  • N-Acetylmuramoyl-L-alanine Amidase