High-molecular-mass penicillin-binding proteins (PBPs) are enzymes that catalyze the biosynthesis of bacterial cell wall peptidoglycan. The Gram-positive bacterial pathogen
Streptococcus agalactiae
(group B
streptococcus
, or GBS) produces five high-molecular-mass PBPs, namely, PBP1A, PBP1B, PBP2A, PBP2B, and PBP2X. Among these, only PBP2X is essential for cell viability, whereas the other four PBPs are individually dispensable. The biological function of the four non-essential PBPs is poorly characterized in GBS. We deleted the
pbp1a
,
pbp1b
,
pbp2a
, and
pbp2b
genes individually from a genetically well-characterized serotype V GBS strain, and studied the phenotypes of the four isogenic mutant strains. Compared to the wild-type parental strain (i) none of the
pbp
isogenic mutant strains had a significant growth defect in THY rich medium, (ii) isogenic mutant strains Δ
pbp1a
and Δ
pbp1b
had significantly increased susceptibility to penicillin and ampicillin, and (iii) isogenic mutant strains Δ
pbp1a
and Δ
pbp2b
had significantly longer chain lengths. Using saturated transposon mutagenesis and transposon insertion site sequencing, we determined genes essential for the viability of wild-type GBS strain and each of the four isogenic
pbp
deletion mutant strains in THY rich medium. The
pbp1a
gene is essential for cell viability in the
pbp2b
deletion background. Reciprocally,
pbp2b
is essential in the
pbp1a
deletion background. Moreover, the gene encoding RodA, a peptidoglycan polymerase that works in conjunction with PBP2B, is also essential in the
pbp1a
deletion background. Together, our results suggest functional overlap between PBP1A and PBP2B-RodA complex in GBS cell wall peptidoglycan biosynthesis.
IMPORTANCE
High-molecular-mass penicillin-binding proteins (HMM-PBPs) are enzymes required for bacterial cell-wall biosynthesis. Bacterial pathogen group B
streptococcus
(GBS) produces five distinct HMM-PBPs. The biological functions of these proteins are not well characterized in GBS. In this study, we performed a comprehensive deletion analysis of genes encoding HMM-PBPs in GBS. We found that deleting certain PBP-encoding genes altered bacterial susceptibility to beta-lactam antibiotics, cell morphology, and the essentiality of other enzymes involved in cell-wall peptidoglycan synthesis. The results of our study shed new light on the biological functions of PBPs in GBS.