Cryo-EM structures of the pore-forming A subunit from the Yersinia entomophaga ABC toxin

Piper, Sarah; Brillault, Lou; Rothnagel, Rosalba; Croll, Tristan I.; Box, J.K.; Chassagnon, Irène R.; Scherer, Sebastian; Goldie, Kenneth N.; Jones, Sandra A.; Schepers, Femke; Hartley-Tassell, Lauren E.; Ve, Thomas; Busby, J.N.; Dalziel, Julie E.; Lott, J.S.; Hankamer, Ben; Stahlberg, Henning; Hurst, Mark R. H.; Landsberg, Michael J.

doi:10.1038/s41467-019-09890-8

Cited by 43 publications

(56 citation statements)

References 61 publications

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“…In a previous glycan array screen with the Tc toxin Yen-Tc from the insect pathogen Yersinia entomophaga , 72 out of 432 glycans were identified as potential toxin receptors, amongst them monosaccharides, fucosylated glycans, mannose derivatives, glucose derivatives, sialylated derivatives, glycosaminoglycans, complex N -glycans and derivatives with terminal GlcNAc, GalNAc or galactose 19 . In contrast, Xn-XptA1, Mm-TcdA4 and Yp-TcaATcaB only interacted strongly with heparins/HS (11 out of 36 heparins/HS on the microarray).…”

Section: Discussionmentioning

confidence: 99%

“…It was proposed that the chitinases cause the degradation of the peritrophic membrane within the midgut of insects to facilitate toxin entry 18 . The association of YenTcA with target cells potentially occurs via glycan structures, which were recently identified in a binding screen to associate with both chitinases and directly with YenTc 19 .…”

Section: Introductionmentioning

confidence: 99%

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Glycan-dependent two-step cell adhesion mechanism of Tc toxins

Roderer

Broecker

Sitsel

et al. 2019

Preprint

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15 Toxin complex (Tc) toxins are virulence factors widespread in insect and human 16 bacterial pathogens. Tcs are composed of three subunits: TcA, TcB and TcC. TcA 17 facilitates receptor-toxin interaction and membrane permeation, TcB and TcC 18 form a toxin-encapsulating cocoon. While the mechanisms of holotoxin assembly 19 and prepore-to-pore transition have been well-described, little is known about 20 receptor binding and cellular uptake of Tcs. Here, we identify two classes of 21 glycans, heparins/heparan sulfates and Lewis antigens, that act as receptors for 22 different TcAs from insect-and human pathogenic bacteria. Glycan array 23 screening and electron cryo microscopy (cryo-EM) structures reveal that all tested 24 TcAs bind unexpectedly with their a-helical part of the shell domain to negatively 25 charged heparins. In addition, TcdA1 from the insect-pathogen Photorhabdus 26 luminescens binds to Lewis antigens with micromolar affinity. A cryo-EM 27 structure of the TcdA1-Lewis X complex reveals that the glycan interacts with the 28 receptor-binding domain D of the toxin. Our results suggest a two-step association 29 mechanism of Tc toxins involving glycans on the surface of host cells. 30 31 32

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Glycan-dependent two-step cell adhesion mechanism of Tc toxins

Roderer

Broecker

Sitsel

et al. 2019

Preprint

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“…The key virulence determinant of MH96 is an insecticidal toxin complex (TC) called Yen-TC, which facilitates ingress of MH96 through the insect midgut (Hurst et al, 2011, 2014; Marshall et al, 2012) and is constitutively produced and secreted by MH96 when grown in broth culture at temperatures of 25 °C or lower (Hurst et al, 2011). In addition to the orally active insecticidal activities of Yen-TC, structural characterization and understanding the molecular mechanisms of pore-formation and translocation of cytotoxin has been the focus of much research (Busby et al, 2012, 2013; Landsberg et al, 2011; Piper et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

In vivotranscriptome analysis provides insights into host-dependent expression of virulence factors byYersinia entomophagaMH96, during infection ofGalleria mellonella

Paulson

O’Callaghan

Zhang³

et al. 2020

Preprint

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The function of microbes can be inferred from knowledge of genes specifically expressed in natural environments. Here we report the in vivo transcriptome of the entomopathogenic bacterium Yersinia entomophaga MH96, captured during three stages of intrahemocoelic infection in Galleria mellonella. A total of 1,285 genes were significantly upregulated by MH96 during infection; 829 genes responded to in vivo conditions during at least one stage of infection, 289 responded during two stages of infection and 167 transcripts responded throughout all three stages of infection. Genes upregulated during the earliest infection stage included components of the insecticidal toxin complex Yen-TC (chi1, chi2 and yenC1), genes for rearrangement hotspot element containing protein yenC3, cytolethal distending toxin cdtAB and vegetative insecticidal toxin vip2. Genes more highly expressed throughout the infection cycle included the putative heat-stable enterotoxin yenT and three adhesins (usher-chaperone fimbria, filamentous hemagglutinin and an AidA-like secreted adhesin). Clustering and functional enrichment of gene expression data also revealed expression of genes encoding type III and VI secretion system-associated effectors. Together these data provide insight into the pathobiology of MH96 and serve as an important resource supporting efforts to identify novel insecticidal agents.

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“…The Yen-Tc comprises five major subunit proteins, Tc-A (Yen A1 and YenA2), Tc-B (YenB) and Tc-C (YenC1 and YenC2), and two associated chitinases, Chi1 and Chi2, which are required for full toxicity following ingestion by susceptible insect hosts. 13,14 Currently, a bioassay targeting P. xylostella larvae is the most sensitive method for detecting the Yen-Tc, which has a six days median lethal dose of c. 4 ng per larva. 15 This concentration is below the detection limits of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis with silver staining.…”

Section: Introductionmentioning

confidence: 99%

Assessment of toxicity and persistence of Yersinia entomophaga and its Yen‐Tc associated toxin

Hurst

Jones

Young

et al. 2020

Pest Management Science

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BACKGROUND: The insect-pathogenic bacterium Yersinia entomophaga MH96 is currently under development as a microbial pesticide active against various pasture and crop pests such as the diamondback moth Plutella xylostella and the cotton bollworm Helicoverpa armigeria. To enable nonrestricted field trials of Y. entomophaga MH96, information on the persistence and nontarget effects of the bacterium and its Yen-Tc proteinaceous toxin are required. RESULTS: The Y. entomophaga Yen-Tc associated toxin was found to have limited persistence on foliage and is inactivated by UV light. The Yen-Tc was rapidly degraded in ovine or bovine rumen fluid or the intestinal fluid of H. armigera. In H. armigera an intestinal protein of >50 kDa was found to cleave the Yen-Tc bond. Assessment of Y. entomophaga persistence on foliage and in soil found that after 42 days the bacterium could not be detected in soil at 20% soil moisture content but persisted for 72 days at 30-40% soil moisture. Nontarget effects of Y. entomophaga towards earthworms found that the bacterium afforded no adverse effects on worm growth or behavior. A summary of historic Yen-Tc and Y. entomophaga persistence and toxicity data is presented. CONCLUSION: The bacterium Y. entomophaga and its Yen-Tc associated toxin have limited persistence in the environment, with the Yen-Tc being susceptible to UV inactivation and proteolytic degradation, and the bacterium persisting longer in soil of a high moisture content.

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Cryo-EM structures of the pore-forming A subunit from the Yersinia entomophaga ABC toxin

Cited by 43 publications

References 61 publications

Glycan-dependent two-step cell adhesion mechanism of Tc toxins

Glycan-dependent two-step cell adhesion mechanism of Tc toxins

In vivotranscriptome analysis provides insights into host-dependent expression of virulence factors byYersinia entomophagaMH96, during infection ofGalleria mellonella

Assessment of toxicity and persistence of Yersinia entomophaga and its Yen‐Tc associated toxin

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