Inhibitors of the
            <i>Neisseria meningitidis</i>
            PilF ATPase provoke type IV pilus disassembly

Aubey, Flore; Corre, Jean-Philippe; Kong, Y.; Xu, Ximing; Obino, Dorian; Goussard, Sylvie; Lapeyrere, Catherine; Souphron, Judith; Couturier, Cédric; Renard, Stéphane; Duménil, Gérard

doi:10.1073/pnas.1817757116

Cited by 35 publications

(30 citation statements)

References 29 publications

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“…Targeting the nucleotide-binding site or packing-unit interface to inhibit ATPase activity in T4P-like systems may have therapeutic value as these systems play a major role in virulence for many pathogens. Indeed, there are two recent reports of small-molecule inhibitors of the Neisseria meningitidis T4aP that target pilus polymerization and depolymerization dynamics to reduce virulence 41,42 . One of these drugs targets PilB directly 41 , although whether this drug binds the nucleotide-binding site or packing-unit interface is not yet clear.…”

Section: Discussionmentioning

confidence: 99%

Multiple conformations facilitate PilT function in the type IV pilus

et al. 2019

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Type IV pilus-like systems are protein complexes that polymerize pilin fibres. They are critical for virulence in many bacterial pathogens. Pilin polymerization and depolymerization are powered by motor ATPases of the PilT/VirB11-like family. This family is thought to operate with C2 symmetry; however, most of these ATPases crystallize with either C3 or C6 symmetric conformations. The relevance of these conformations is unclear. Here, we determine the X-ray structures of PilT in four unique conformations and use these structures to classify the conformation of available PilT/VirB11-like family member structures. Single particle electron cryomicroscopy (cryoEM) structures of PilT reveal condition-dependent preferences for C2, C3, and C6 conformations. The physiologic importance of these conformations is validated by coevolution analysis and functional studies of point mutants, identifying a rare gain-of-function mutation that favours the C2 conformation. With these data, we propose a comprehensive model of PilT function with broad implications for PilT/VirB11-like family members.

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Section: Discussionmentioning

confidence: 99%

Multiple conformations facilitate PilT function in the type IV pilus

et al. 2019

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“…Coincidentally, P4MP4 affected N. meningitidis aggregation and T4P in a manner resembling that of phenothiazines despite their different routes of discovery. Biochemical studies in this investigation pointed to PilF, the PilB equivalent in the N. meningitidis T4P system, as the potential target of P4MP4 instead of the Na 1 -NQR complex (30). It should be emphasized that the T4PM was not specifically targeted by the antiaggregation (29) or the antiadhesion (30) assays in either case for the discovery of phenothiazines or P4MP4.…”

mentioning

confidence: 82%

High-Throughput Screen for Inhibitors of the Type IV Pilus Assembly ATPase PilB

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Vogelaar

Sobrado

et al. 2021

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The bacterial type IV pilus (T4P) is a prominent virulence factor in many significant human pathogens, some of which have become increasingly antibiotic resistant. Antivirulence chemotherapeutics are considered a promising alternative to antibiotics because they target the disease process instead of bacterial viability. However, a roadblock to the discovery of anti-T4P compounds is the lack of a high-throughput screen (HTS) that can be implemented relatively easily and economically. Here, we describe the first HTS for the identification of inhibitors specifically against the T4P assembly ATPase PilB in vitro. Chloracidobacterium thermophilum PilB (CtPilB) had been demonstrated to have robust ATPase activity and the ability to bind its expected ligands in vitro. We utilized CtPilB and MANT-ATP, a fluorescent ATP analog, to develop a binding assay and adapted it for an HTS. As a proof of principle, we performed a pilot screen with a small compound library of kinase inhibitors and identified quercetin as a PilB inhibitor in vitro. Using Myxococcus xanthus as a model bacterium, we found quercetin to reduce its T4P-dependent motility and T4P assembly in vivo. These results validated our HTS as effective in identifying PilB inhibitors. This assay may prove valuable in seeking leads for the development of antivirulence chemotherapeutics against PilB, an essential and universal component of all bacterial T4P systems. IMPORTANCE Many bacterial pathogens use their type IV pili (T4P) to facilitate and maintain infection of a human host. Small chemical compounds that inhibit the production or assembly of T4P hold promise in the treatment and prevention of infections, especially in the era of increasing threats from antibiotic-resistant bacteria. However, few chemicals are known to have inhibitory or anti-T4P activity. Their identification has not been easy due to the lack of a method for the screening of compound collections or libraries on a large scale. Here, we report the development of an assay that can be scaled up to screen compound libraries for inhibitors of a critical T4P assembly protein. We further demonstrate that it is feasible to use whole cells to examine potential inhibitors for their activity against T4P assembly in a bacterium.

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“…Two recent studies have identified inhibitors of neisserial tfp. The first inhibitor, referred to as compound B in the publication, was identified through a phenotypic screen and successfully prevented the adherence and formation of N. meningitidis microcolonies on the human umbilical vein endothelial cell surface [ 181 ]. Cellular and in vitro experiments showed that compound B could inhibit the PilF ATPase enzymatic activity resulting in lowered surface expression levels of tfp.…”

Section: Gonococcal Virulence Factors As Targets For Inhibitor Desmentioning

confidence: 99%

“…The effects of both compounds on piliation were fast acting, reflecting the rapid dynamics of the tfp [ 181 , 182 ]. Additionally, both studies showed that although these compounds were initially designed to inhibit N. meningitidis , they are also effective on other Gram-negative tfp-expressing bacterial pathogens such as N. gonorrhoeae and P. aeruginosa .…”

Section: Gonococcal Virulence Factors As Targets For Inhibitor Desmentioning

confidence: 99%

Anti-Virulence Therapeutic Approaches for Neisseria gonorrhoeae

et al. 2021

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While antimicrobial resistance (AMR) is seen in both Neisseria gonorrhoeae and Neisseria meningitidis, the former has become resistant to commonly available over-the-counter antibiotic treatments. It is imperative then to develop new therapies that combat current AMR isolates whilst also circumventing the pathways leading to the development of AMR. This review highlights the growing research interest in developing anti-virulence therapies (AVTs) which are directed towards inhibiting virulence factors to prevent infection. By targeting virulence factors that are not essential for gonococcal survival, it is hypothesized that this will impart a smaller selective pressure for the emergence of resistance in the pathogen and in the microbiome, thus avoiding AMR development to the anti-infective. This review summates the current basis of numerous anti-virulence strategies being explored for N. gonorrhoeae.

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Inhibitors of the Neisseria meningitidis PilF ATPase provoke type IV pilus disassembly

Cited by 35 publications

References 29 publications

Multiple conformations facilitate PilT function in the type IV pilus

Multiple conformations facilitate PilT function in the type IV pilus

High-Throughput Screen for Inhibitors of the Type IV Pilus Assembly ATPase PilB

Anti-Virulence Therapeutic Approaches for Neisseria gonorrhoeae

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