Piericidin A1 Blocks
            <i>Yersinia</i>
            Ysc Type III Secretion System Needle Assembly

Morgan, Joe P.; Duncan, Miles C.; Johnson, Kevin S.; Diepold, Andreas; Lam, Hanh; Dupzyk, Allison J.; Martin, Lexi R.; Wong, Weng Ruh; Armitage, Judith P.; Linington, Roger G.; Auerbuch, Victoria

doi:10.1128/msphere.00030-17

Cited by 20 publications

(22 citation statements)

References 55 publications

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“…The proton motive force is important for the Yersinia flagellar T3SS, as the proton motive force inhibitor carbonyl cyanide m-chlorophenylhydrazone (CCCP) prevents Yersinia motility (33,61). Yet the majority of our compounds do not affect flagellar motility.…”

Section: Figmentioning

confidence: 93%

Synthetic Cyclic Peptomers as Type III Secretion System Inhibitors

Lam

Schwochert

Lao

et al. 2017

Antimicrob Agents Chemother

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Antibiotic-resistant bacteria are an emerging threat to global public health. New classes of antibiotics and tools for antimicrobial discovery are urgently needed. Type III secretion systems (T3SS), which are required by dozens of Gramnegative bacteria for virulence but largely absent from nonpathogenic bacteria, are promising virulence blocker targets. The ability of mammalian cells to recognize the presence of a functional T3SS and trigger NF-B activation provides a rapid and sensitive method for identifying chemical inhibitors of T3SS activity. In this study, we generated a HEK293 stable cell line expressing green fluorescent protein (GFP) driven by a promoter containing NF-B enhancer elements to serve as a readout of T3SS function. We identified a family of synthetic cyclic peptide-peptoid hybrid molecules (peptomers) that exhibited dose-dependent inhibition of T3SS effector secretion in Yersinia pseudotuberculosis and Pseudomonas aeruginosa without affecting bacterial growth or motility. Among these inhibitors, EpD-3=N, EpD-1,2N, EpD-1,3=N, EpD-1,2,3=N, and EpD-1,2,4=N exhibited strong inhibitory effects on translocation of the Yersinia YopM effector protein into mammalian cells (Ͼ40% translocation inhibition at 7.5 M) and showed no toxicity to mammalian cells at 240 M. In addition, EpD-3=N and EpD-1,2,4=N reduced the rounding of HeLa cells caused by the activity of Yersinia effector proteins that target the actin cytoskeleton. In summary, we have discovered a family of novel cyclic peptomers that inhibit the injectisome T3SS but not the flagellar T3SS.

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Section: Figmentioning

confidence: 93%

Synthetic Cyclic Peptomers as Type III Secretion System Inhibitors

Lam

Schwochert

Lao

et al. 2017

Antimicrob Agents Chemother

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“…The T3SS transports Yops to the cytoplasm of host cells through a syringe-like system, formed mainly by YscF and translocator Yops [ 66 , 68 ]. YopB and YopD, two similar hydrophobic proteins referred to as translocators, form a multimeric integral membrane complex in the membrane of eukaryotic cells [ 69 ].…”

Section: Pathogenesis Of Y Enterocolitica Infementioning

confidence: 99%

The Most Important Virulence Markers of Yersinia enterocolitica and Their Role during Infection

Bancerz‐Kisiel

Pieczywek

Łada

et al. 2018

Genes

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Yersinia enterocolitica is the causative agent of yersiniosis, a zoonotic disease of growing epidemiological importance with significant consequences for public health. This pathogenic species has been intensively studied for many years. Six biotypes (1A, 1B, 2, 3, 4, 5) and more than 70 serotypes of Y. enterocolitica have been identified to date. The biotypes of Y. enterocolitica are divided according to their pathogenic properties: the non-pathogenic biotype 1A, weakly pathogenic biotypes 2–5, and the highly pathogenic biotype 1B. Due to the complex pathogenesis of yersiniosis, further research is needed to expand our knowledge of the molecular mechanisms involved in the infection process and the clinical course of the disease. Many factors, both plasmid and chromosomal, significantly influence these processes. The aim of this study was to present the most important virulence markers of Y. enterocolitica and their role during infection.

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“…Compound 20 (CAS# 489402-27-9) was obtained from TimTek. The anti-YscF antibody was raised in rabbits against the Y. pseudotuberculosis YscF peptide KDKPDNPALLADLQH (Morgan et al, 2017 ). DMSO concentration did not exceed 0.2%, except where indicated.…”

Section: Methodsmentioning

confidence: 99%

“…Quantification of YscF staining on the bacterial surface was carried out as described previously (Morgan et al, 2017 ). Indicated concentrations of compounds, or the equivalent volumes of DMSO, were added to 26°C-grown low calcium medium cultures and treated cultures were shifted to 37°C for 3 h. Bacteria were fixed by the addition of a mix of 800 μl 4% paraformaldehyde, 1 μl 25% glutaraldehyde, and 40 μl 0.5 M NaPO 4 pH 7.4 to 500 μl of bacterial culture for 15 min at room temperature followed by 30 min or longer on ice.…”

Section: Methodsmentioning

confidence: 99%

An Experimental Pipeline for Initial Characterization of Bacterial Type III Secretion System Inhibitor Mode of Action Using Enteropathogenic Yersinia

Morgan

Lam

Delgado

et al. 2018

Front. Cell. Infect. Microbiol.

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Dozens of Gram negative pathogens use one or more type III secretion systems (T3SS) to disarm host defenses or occupy a beneficial niche during infection of a host organism. While the T3SS represents an attractive drug target and dozens of compounds with T3SS inhibitory activity have been identified, few T3SS inhibitors have been validated and mode of action determined. One issue is the lack of standardized orthogonal assays following high throughput screening. Using a training set of commercially available compounds previously shown to possess T3SS inhibitory activity, we demonstrate the utility of an experiment pipeline comprised of six distinct assays to assess the stages of type III secretion impacted: T3SS gene copy number, T3SS gene expression, T3SS basal body and needle assembly, secretion of cargo through the T3SS, and translocation of T3SS effector proteins into host cells. We used enteropathogenic Yersinia as the workhorse T3SS-expressing model organisms for this experimental pipeline, as Yersinia is sensitive to all T3SS inhibitors we tested, including those active against other T3SS-expressing pathogens. We find that this experimental pipeline is capable of rapidly distinguishing between T3SS inhibitors that interrupt the process of type III secretion at different points in T3SS assembly and function. For example, our data suggests that Compound 3, a malic diamide, blocks either activity of the assembled T3SS or alters the structure of the T3SS in a way that blocks T3SS cargo secretion but not antibody recognition of the T3SS needle. In contrast, our data predicts that Compound 4, a haloid-containing sulfonamidobenzamide, disrupts T3SS needle subunit secretion or assembly. Furthermore, we suggest that misregulation of copy number control of the pYV virulence plasmid, which encodes the Yersinia T3SS, should be considered as a possible mode of action for compounds with T3SS inhibitory activity against Yersinia.

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Piericidin A1 Blocks Yersinia Ysc Type III Secretion System Needle Assembly

Cited by 20 publications

References 55 publications

Synthetic Cyclic Peptomers as Type III Secretion System Inhibitors

Synthetic Cyclic Peptomers as Type III Secretion System Inhibitors

The Most Important Virulence Markers of Yersinia enterocolitica and Their Role during Infection

An Experimental Pipeline for Initial Characterization of Bacterial Type III Secretion System Inhibitor Mode of Action Using Enteropathogenic Yersinia

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