title: 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.
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.
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, IN, 47405, USA.
authors.forename: Ho-Ching Tiffany
authors.initials: HC
authors.lastname: Tsui
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, IN, 47405, USA.
authors.forename: Jiaqi J
authors.initials: JJ
authors.lastname: Zheng
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, IN, 47405, USA.
authors.forename: Ariel N
authors.initials: AN
authors.lastname: Magallon
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, IN, 47405, USA.
authors.forename: John D
authors.initials: JD
authors.lastname: Ryan
authors.affiliation: Department of Microbiology and Immunology, Harvard Medical School, Boston, MA, 02115, USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Yunck
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, IN, 47405, USA.
authors.forename: Britta E
authors.initials: BE
authors.lastname: Rued
authors.affiliation: Department of Microbiology and Immunology, Harvard Medical School, Boston, MA, 02115, USA.
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Bernhardt
authors.affiliation: Department of Biology, Indiana University Bloomington, Bloomington, IN, 47405, USA.
authors.forename: Malcolm E
authors.initials: ME
authors.lastname: Winkler
mesh_terms: D019881:Aminoacyltransferases / Q000235:genetics* / Q000378:metabolism; D016695:Glycosyltransferases / Q000378:metabolism; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000378:metabolism; D046915:Penicillin-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D010457:Peptidoglycan / Q000096:biosynthesis* / Q000378:metabolism; D010458:Peptidyl Transferases / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D013296:Streptococcus pneumoniae / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism
chemical_list: D046915:Penicillin-Binding Proteins; D010457:Peptidoglycan; D019881:Aminoacyltransferases; C478701:penicillin-binding protein 2b, Streptococcus; D010458:Peptidyl Transferases; D016695:Glycosyltransferases; D009238:N-Acetylmuramoyl-L-alanine Amidase