Moenomycin Resistance Mutations in <i>Staphylococcus aureus</i> Reduce Peptidoglycan Chain Length and Cause Aberrant Cell Division

Rebets, Yuriy; Lupoli, Tania J.; Qiao, Yuan; Schirner, Kathrin; Villet, Régis; Hooper, David C.; Kahne, Daniel; Walker, Suzanne

doi:10.1021/cb4006744

Cited by 52 publications

(62 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To obtain short peptidoglycan oligomers, we incubated synthetic Lipid II with a mutant form of S. aureus SgtB, a monofunctional peptidoglycan glycosyltransferase that polymerizes Lipid II. The mutant, SgtB*, contains a Y181D amino acid substitution in the active site cleft where the elongating polymer binds 33 , and as a result products are released prematurely (Fig. 2b).…”

Section: Resultsmentioning

confidence: 99%

In vitro reconstitution demonstrates the cell wall ligase activity of LCP proteins

et al. 2017

Self Cite

View full text Add to dashboard Cite

Sacculus is a peptidoglycan matrix that protects bacteria from osmotic lysis. In Gram-positive organisms, the sacculus is densely functionalized with glycopolymers important for survival, but how assembly occurs is not known. In Staphylococcus aureus, three LCP family members have been implicated in attaching the major glycopolymer wall teichoic acid (WTA) to peptidoglycan, but ligase activity has not been demonstrated for these or any other LCP proteins. Using WTA and peptidoglycan substrates produced chemoenzymatically, we show that all three proteins can transfer WTA precursors to nascent peptidoglycan, establishing that LCP proteins are peptidoglycan-glycopolymer ligases. Although all S. aureus LCP proteins have the capacity to attach WTA to PG, we show that their cellular functions are not redundant. Strains lacking lcpA have phenotypes similar to WTA null strains, indicating that this is the most important WTA ligase. This work provides a foundation for studying how LCP enzymes participate in cell wall assembly.

show abstract

Section: Resultsmentioning

confidence: 99%

In vitro reconstitution demonstrates the cell wall ligase activity of LCP proteins

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…As part of the cell wall stimulon 22 , mgt is positively regulated by cell wall stress and participates in the polymerization of lipid II into nascent peptidoglycan 23 . Recent work has shown that mgt mutations cause peptidoglycan chain length reduction as well as alterations in cellular morphology and division site placement 24 .…”

Section: Resultsmentioning

confidence: 99%

Genomic signatures of experimental adaptation to antimicrobial peptides in Staphylococcus aureus

Johnston

Dobson

Rolff

2015

Preprint

View full text Add to dashboard Cite

ObjectivesThe evolution of resistance against antimicrobial peptides has long been considered unlikely due to their mechanism of action, yet experimental selection with AMPs results in rapid evolution of resistance in several species of bacteria. Although numerous studies have utilized mutant screens to identify loci that determine AMP susceptibility, there is a dearth of data concerning the genomic changes which accompany experimental evolution of AMP resistance.MethodsUsing genome re-sequencing we analysed the mutations which arise during experimental evolution of resistance to the cationic AMPs iseganan, melittin and pexiganan, as well as to a combination of melittin and pexiganan, or to the aminoglycoside antibiotic streptomycin.ResultsAnalysis of 17 independently replicated Staphylococcus aureus selection lines, including unselected controls, showed that each AMP selected for mutations at distinct loci. We identify mutations in genes involved in the synthesis and maintenance of the cell envelope. This includes genes previously identified from mutant screens for AMP resistance, and genes involved in the response to AMPs and cell-wall-active antibiotics. Furthermore, transposon insertion mutants were used to verify that a number of the identified genes are directly involved in determining AMP susceptibility.ConclusionsStrains selected for AMP resistance under controlled experimental evolution displayed consistent AMP-specific mutations in genes which determine AMP susceptibility. This suggests that different routes to evolve resistance are favored within a controlled genetic background.

show abstract

“…To compare the stem peptide preferences of Ef PBPX, Sg PBPX, and Sa PBP4, we made uncrosslinked peptidoglycan oligomers from the six Lipid II variants by incubating them with SgtB Y181D (SgtB*), a peptidoglycan polymerase that makes short peptidoglycan chains. 12 BDL and PBPs were then added and the reaction products analyzed by western blot. Sa PBP4 labeled all the substrates, although four-fold higher concentrations of 4 – 6 were required to produce enough labeled material to detect (Figure S7).…”

mentioning

confidence: 99%

Identification of a Functionally Unique Family of Penicillin-Binding Proteins

et al. 2017

Self Cite

View full text Add to dashboard Cite

Penicillin-binding proteins (PBPs) are enzymes involved in the assembly of the bacterial cell wall, a major target for antibiotics. These proteins are classified by mass into high molecular weight PBPs, which are transpeptidases that form peptidoglycan crosslinks, and low molecular weight PBPs, which are typically hydrolases. We report a functionally unique family of low molecular weight PBPs that act as transpeptidases rather than hydrolases, but they do not crosslink peptidoglycan. We show that these PBPs can exchange D-amino acids bearing chemical tags or affinity handles into peptidoglycan precursors, including Lipid II, enabling biochemical studies of proteins involved in cell wall assembly. We report that, in two organisms, the PBPs incorporate lysine into peptidoglycan and, further, that this linkage can occur via attack of the primary ε-amine side chain, an unprecedented reaction for a PBP.

show abstract

Moenomycin Resistance Mutations in Staphylococcus aureus Reduce Peptidoglycan Chain Length and Cause Aberrant Cell Division

Cited by 52 publications

References 53 publications

In vitro reconstitution demonstrates the cell wall ligase activity of LCP proteins

In vitro reconstitution demonstrates the cell wall ligase activity of LCP proteins

Genomic signatures of experimental adaptation to antimicrobial peptides in Staphylococcus aureus

Identification of a Functionally Unique Family of Penicillin-Binding Proteins

Contact Info

Product

Resources

About