Coupling of polymerase and carrier lipid phosphatase prevents product inhibition in peptidoglycan synthesis

Hernández-Rocamora, Víctor M.; Otten, Christian; Radkov, Atanas; Simorre, Jean-Pierre; Breukink, Eefjan; VanNieuwenhze, Michael S.; Vollmer, Waldemar

doi:10.1016/j.tcsw.2018.04.002

Cited by 23 publications

(21 citation statements)

References 43 publications

(69 reference statements)

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“…PBP1B interacts with a number of proteins including PBP3 (ref. 7 ), FtsW 13 , FtsN 20 , LpoB 40 , 57 , 58 , CpoB 59 , MipA 60 , PgpB 61 and now ZipA. It is unlikely that all PBP1B molecules in the cell interact with all of these proteins at all times.…”

Section: Discussionmentioning

confidence: 99%

Z-ring membrane anchors associate with cell wall synthases to initiate bacterial cell division

et al. 2018

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During the transition from elongation to septation, Escherichia coli establishes a ring-like peptidoglycan growth zone at the future division site. This preseptal peptidoglycan synthesis does not require the cell division-specific peptidoglycan transpeptidase PBP3 or most of the other cell division proteins, but it does require FtsZ, its membrane-anchor ZipA and at least one of the bi-functional transglycosylase-transpeptidases, PBP1A or PBP1B. Here we show that PBP1A and PBP1B interact with ZipA and localise to preseptal sites in cells with inhibited PBP3. ZipA stimulates the glycosyltransferase activity of PBP1A. The membrane-anchored cell division protein FtsN localises at preseptal sites and stimulates both activities of PBP1B. Genes zipA and ftsN can be individually deleted in ftsA* mutant cells, but the simultaneous depletion of both proteins is lethal and cells do not establish preseptal sites. Our data support a model according to which ZipA and FtsN-FtsA have semi-redundant roles in connecting the cytosolic FtsZ ring with the membrane-anchored peptidoglycan synthases during the preseptal phase of envelope growth.

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Section: Discussionmentioning

confidence: 99%

Z-ring membrane anchors associate with cell wall synthases to initiate bacterial cell division

et al. 2018

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“…The PBP1A and PBP1B play major roles in peptidoglycan polymerization and their simultaneous inactivation is lethal 24 . Recently, an interaction between PgpB and PBP1B was shown, highlighting a coupling event between peptidoglycan polymerization and C 55 -PP recycling 25 . We then addressed whether a strict interdependency exists between these major PBPs and C 55 -PP phosphatases.…”

Section: Multiple Pap2 Knockout Mutants Are Sensitive To Small Hydropmentioning

confidence: 99%

Insight into the dual function of lipid phosphate phosphatase PgpB involved in two essential cell-envelope metabolic pathways in Escherichia coli

Tian

Auger

Manat

et al. 2020

Sci Rep

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Ubiquitous PAP2 lipid phosphatases are involved in a wide array of central physiological functions. PgpB from Escherichia coli constitutes the archetype of this subfamily of membrane proteins. It displays a dual function by catalyzing the biosynthesis of two essential lipids, the phosphatidylglycerol (PG) and the undecaprenyl phosphate (C 55-P). C 55-P constitutes a lipid carrier allowing the translocation of peptidoglycan subunits across the plasma membrane. PG and C 55-p are synthesized in a redundant manner by PgpB and other PAP2 and/or unrelated membrane phosphatases. Here, we show that PgpB is the sole, among these multiple phosphatases, displaying this dual activity. The inactivation of PgpB does not confer any apparent growth defect, but its inactivation together with another PAP2 alters the cell envelope integrity increasing the susceptibility to small hydrophobic compounds. Evidence is also provided of an interplay between PAP2s and the peptidoglycan polymerase PBP1A. In contrast to PGP hydrolysis, which relies on a His/Asp/His catalytic triad of PgpB, the mechanism of C 55-PP hydrolysis appeared as only requiring the His/Asp diad, which led us to hypothesize distinct processes. Moreover, thermal stability analyses highlighted a substantial structural change upon phosphate binding by PgpB, supporting an induced-fit model of action. Undecaprenyl phosphate (C 55-P) plays an essential role in the biogenesis of bacterial envelope polysaccharides such as the peptidoglycan 1. It is used as a lipid carrier allowing the translocation of polymer subunits across the plasma membrane to the outer site, where the polymers are assembled 2 (Supplementary Fig. S1). The synthesis of C 55-P proceeds via the hydrolysis of its precursor, the undecaprenyl pyrophosphate (C 55-PP), itself being de novo synthesized at the cytosolic side of the plasma membrane or released during subunits polymerization at the outer side (Supplementary Fig. S1) 1. Four integral membrane enzymes catalyzing C 55-PP hydrolysis have been identified in Escherichia coli: BacA, PgpB, YbjG and LpxT (formerly YeiU), which belong to two unrelated protein families: BacA and Phosphatidic Acid Phosphatases of type 2 (PAP2) 3-5. None of these enzymes is essential for growth, but the simultaneous knockout of bacA, pgpB and ybjG genes elicits a lethal phenotype due to a default of C 55-P supply 4. These enzymes have their active sites oriented towards the periplasm, suggesting they may rather be involved in C 55-PP recycling 5-10. The mechanism of translocation of C 55-P back to the inner side of the membrane is yet unknown. The structure of BacA, which is reminiscent to that of transporters, raised the hypothesis that it may also catalyze the flip of C 55-P 7,10. The lack of a known cytoplasm-oriented C 55-PP phosphatase raised also the question as to whether BacA and PAP2s are required for the dephosphorylation of de novo synthesized C 55-PP (Supplementary Fig. S1). PgpB is involved in another essential metabolic pathway, i.e. the synthesis of phosphatidylglyc...

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“…For example, E. coli cells induced to grow with low Und-P readily lyse in media containing aztreonam or mecillinam, β-lactam antibiotics that inhibit the PG synthesizing activity of penicillin binding proteins (PBPs) (Jorgenson et al, 2019). Similarly, deleting pgpB sensitizes cells to cefsulodin, another β-lactam antibiotic (Hernandez-Rocamora et al, 2018). β-lactam antibiotics also synergize with the UppS inhibitor (and fertility drug) clomiphene to kill methicillinresistant Staphylococcus aureus (Farha et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

A genetic screen to identify factors affected by undecaprenyl phosphate recycling uncovers novel connections to morphogenesis inEscherichia coli

Jorgenson

Bryant

2020

Preprint

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Undecaprenyl phosphate (Und-P) is an essential lipid carrier that ferries cell wall intermediates across the cytoplasmic membrane in bacteria. Und-P is generated by dephosphorylating undecaprenyl diphosphate (Und-PP). In Escherichia coli, BacA, PgpB, YbjG, and LpxT dephosphorylate Und-PP and are conditionally essential. To identify vulnerabilities that arise when Und-P metabolism is defective, we developed a genetic screen for synthetic mutations in combination with ΔybjG ΔlpxT ΔbacA. The screen uncovered system-wide connections, including novel connections to cell division, DNA replication and repair, signal transduction, and glutathione metabolism. Further analysis revealed several new morphogenes; loss of one of these, qseC, caused cells to enlarge and lyse. QseC is the sensor kinase component of the QseBC two-component system. In the absence of QseC, the QseB response regulator is overactivated by PmrB cross-phosphorylation. Here, we show that deleting qseB completely reverses the shape defect of ΔqseC cells, as does overexpressing rprA (a small RNA). Surprisingly, deleting pmrB only partially suppressed qseC-related shape defects. Thus, QseB is activated by multiple factors in the absence of QseC and functions ascribed to QseBC may be related to cell wall defects. Altogether, our findings provide a framework for identifying new determinants of cell integrity that could be targeted in future therapies.

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Coupling of polymerase and carrier lipid phosphatase prevents product inhibition in peptidoglycan synthesis

Cited by 23 publications

References 43 publications

Z-ring membrane anchors associate with cell wall synthases to initiate bacterial cell division

Z-ring membrane anchors associate with cell wall synthases to initiate bacterial cell division

Insight into the dual function of lipid phosphate phosphatase PgpB involved in two essential cell-envelope metabolic pathways in Escherichia coli

A genetic screen to identify factors affected by undecaprenyl phosphate recycling uncovers novel connections to morphogenesis inEscherichia coli

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