The Lantibiotic NAI-107 Binds to Bactoprenol-bound Cell Wall Precursors and Impairs Membrane Functions

Münch, Daniela; Müller, Anna; Schneider, Tanja; Kohl, Bastian; Wenzel, Michaela; Bandow, Julia E.; Maffioli, Sonia; Sosio, Margherita; Donadio, Stefano; Wimmer, Reinhard; Sahl, Hans‐Georg

doi:10.1074/jbc.m113.537449

Cited by 76 publications

(70 citation statements)

References 56 publications

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“…This is similar to what was observed in the UppS depletion strain (16,46) and a strain depleted of lipid II flippases (5). The lantibiotic NAI-107 (which targets UPP-linked cell wall precursors like lipids I and II and precursors of WTA) leads to a similar terminal phenotype (47).…”

Section: Figsupporting

confidence: 67%

Depletion of Undecaprenyl Pyrophosphate Phosphatases Disrupts Cell Envelope Biogenesis in Bacillus subtilis

et al. 2016

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The integrity of the bacterial cell envelope is essential to sustain life by countering the high turgor pressure of the cell and providing a barrier against chemical insults. In Bacillus subtilis, synthesis of both peptidoglycan and wall teichoic acids requires a common C 55 lipid carrier, undecaprenyl-pyrophosphate (UPP), to ferry precursors across the cytoplasmic membrane. The synthesis and recycling of UPP requires a phosphatase to generate the monophosphate form Und-P, which is the substrate for peptidoglycan and wall teichoic acid synthases. Using an optimized clustered regularly interspaced short palindromic repeat (CRISPR) system with catalytically inactive ("dead") CRISPR-associated protein 9 (dCas9)-based transcriptional repression system (CRISPR interference [CRISPRi]), we demonstrate that B. subtilis requires either of two UPP phosphatases, UppP or BcrC, for viability. We show that a third predicted lipid phosphatase (YodM), with homology to diacylglycerol pyrophosphatases, can also support growth when overexpressed. Depletion of UPP phosphatase activity leads to morphological defects consistent with a failure of cell envelope synthesis and strongly activates the M -dependent cell envelope stress response, including bcrC, which encodes one of the two UPP phosphatases. These results highlight the utility of an optimized CRISPRi system for the investigation of synthetic lethal gene pairs, clarify the nature of the B. subtilis UPP-Pase enzymes, and provide further evidence linking the M regulon to cell envelope homeostasis pathways. IMPORTANCEThe emergence of antibiotic resistance among bacterial pathogens is of critical concern and motivates efforts to develop new therapeutics and increase the utility of those already in use. The lipid II cycle is one of the most frequently targeted processes for antibiotics and has been intensively studied. Despite these efforts, some steps have remained poorly defined, partly due to genetic redundancy. CRISPRi provides a powerful tool to investigate the functions of essential genes and sets of genes. Here, we used an optimized CRISPRi system to demonstrate functional redundancy of two UPP phosphatases that are required for the conversion of the initially synthesized UPP lipid carrier to Und-P, the substrate for the synthesis of the initial lipid-linked precursors in peptidoglycan and wall teichoic acid synthesis.

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Section: Figsupporting

confidence: 67%

Depletion of Undecaprenyl Pyrophosphate Phosphatases Disrupts Cell Envelope Biogenesis in Bacillus subtilis

et al. 2016

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“…The result of binding to and sequestering lipid II is inhibition of bacterial cell wall biosynthesis at the transglycosylation step. There are additional proposed mechanisms of action after lipid II binding, which include membrane interaction with or without pore formation (12,40,41). Recently, a candidate lantibiotic, NAI-107 (microbisporicin), emerged from a screening program designed to find new bacterial cell wall inhibitors (7,13).…”

Section: Figmentioning

confidence: 99%

“…The mechanism of action, as deduced in the studies of the prototype lantibiotic, nisin, is inhibition of the growth of Gram-positive bacteria by interfering with peptidoglycan synthesis by binding to lipid II (8)(9)(10)(11)(12). This novel mechanistic site is distinct from that of other antibiotics, and cross-resistance has not been described (13,14).…”

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confidence: 99%

In Vivo Pharmacokinetics and Pharmacodynamics of the Lantibiotic NAI-107 in a Neutropenic Murine Thigh Infection Model

Lepak

Marchillo

Craig

et al. 2015

Antimicrob Agents Chemother

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“…In contrast to nisin, gallidermin does not usually form pores in the membranes of most bacteria, so an additional mechanism must therefore be responsible for its high effectiveness (51). This mechanism might provoke membrane perturbations, as has been described for NAI-107, as well as other cAMPs (52)(53)(54). From a structural point of view, NAI-107 shares the AviCys of gallidermin and also the lipid II binding pyrophosphate cage structure but is neutrally charged.…”

Section: Discussionmentioning

confidence: 99%

VraH Is the Third Component of the Staphylococcus aureus VraDEH System Involved in Gallidermin and Daptomycin Resistance and Pathogenicity

Popella

Krauss

Ebner

et al. 2016

Antimicrob Agents Chemother

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In bacteria, extracellular signals are transduced into the cell predominantly by two-component systems (TCSs) comprising a regulatory unit triggered by a specific signal. Some of the TCSs control executing units such as ABC transporters involved in antibiotic resistance. For instance, in Staphylococcus aureus, activation of BraSR leads to the upregulation of vraDE expression that encodes an ABC transporter playing a role in bacitracin and nisin resistance. In this study, we show that the small staphylococcal transmembrane protein VraH forms, together with VraDE, a three-component system. Although the expression of vraH in the absence of vraDE was sufficient to mediate low-level resistance, only this VraDEH entity conferred high-level resistance against daptomycin and gallidermin. In most staphylococcal genomes, vraH is located immediately downstream of vraDE, forming an operon, whereas in some species it is localized differently. In an invertebrate infection model, VraDEH significantly enhanced S. aureus pathogenicity. In analogy to the TCS connectors, VraH can be regarded as an ABC connector that modulates the activity of ABC transporters involved in antibiotic resistance.

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The Lantibiotic NAI-107 Binds to Bactoprenol-bound Cell Wall Precursors and Impairs Membrane Functions

Cited by 76 publications

References 56 publications

Depletion of Undecaprenyl Pyrophosphate Phosphatases Disrupts Cell Envelope Biogenesis in Bacillus subtilis

Depletion of Undecaprenyl Pyrophosphate Phosphatases Disrupts Cell Envelope Biogenesis in Bacillus subtilis

In Vivo Pharmacokinetics and Pharmacodynamics of the Lantibiotic NAI-107 in a Neutropenic Murine Thigh Infection Model

VraH Is the Third Component of the Staphylococcus aureus VraDEH System Involved in Gallidermin and Daptomycin Resistance and Pathogenicity

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