The Trimeric Autotransporter Adhesin YadA of Yersinia enterocolitica Serotype O:9 Binds Glycan Moieties

Meuskens, Ina; Leva-Bueno, Juan; Millner, Paul A.; Schütz, Monika; Peyman, Sally A.; Linke, Dirk

doi:10.3389/fmicb.2021.738818

Cited by 9 publications

(15 citation statements)

References 48 publications

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“…RadA was so named because it contains several domains present in YadA, an autotransporter adhesin common in Yersinia spp., including a coiled stalk, head domain, and anchor (42,43). YadA, a virulence factor in Y. enterocolitica, mediates interactions between the bacterium and host cell surface glycans, and promotes autoaggregation (cell to cell attachment) (43)(44)(45). Its similarity to YadA suggests that RadA is an adhesin of the same class and might be secreted by the type V secretion system (19).…”

Section: Discussionmentioning

confidence: 99%

Cell-density regulated adhesins contribute to early disease development and adhesion inRalstonia solanacearum

Carter

Khokhani

Allen

2022

Preprint

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Adhesins (adhesive proteins) help bacteria stick to and colonize diverse surfaces and often contribute to virulence. The genome of the bacterial wilt pathogenRalstonia solanacearum(Rs) encodes dozens of putative adhesins, some of which are upregulated during plant pathogenesis. Little is known about the role of these proteins in bacterial wilt disease. During tomato colonization, three putative Rs adhesin genes were upregulated in a ΔphcAquorum sensing mutant that cannot respond to high cell densities:radA(Ralstonia adhesin),rcpA(Ralstonia collagen-like protein), andrcpB. Based on this differential gene expression, we hypothesized that adhesins negatively regulated by PhcA contribute to early disease stages when Rs experiences a low cell density. During root colonizationRsupregulatedrcpAandrcpBrelative to bacteria in the stem at mid-disease, but notradA. Root attachment assays and confocal microscopy withΔrcpA/BandΔradArevealed that all three adhesins helpRsattach to tomato seedling roots. Biofilm assays on abiotic surfaces found thatRsdoes not require RadA, RcpA, or RcpB for interbacterial attachment (cohesion), but these proteins are essential for anchoring aggregates to a surface (adhesion). However,Rsdid not require the adhesins for later disease stagesin planta, including colonization of the root endosphere and stems. Interestingly, all three adhesins were essential for full competitive fitnessin planta. Together, these infection stage-specific assays identified three proteins that contribute to adhesion and the critical first host-pathogen interaction in bacterial wilt disease.

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Section: Discussionmentioning

confidence: 99%

Cell-density regulated adhesins contribute to early disease development and adhesion inRalstonia solanacearum

Carter

Khokhani

Allen

2022

Preprint

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“…First, an existing pASK‐Iba2‐Sumo construct (I. Meuskens, P. E. Kristiansen, B. Bardiaux, V. R. Koynarev, D. Hatlem, K. Prydz, R. Lund, N. Izadi‐Pruneyre, D. Linke) was linearized by PCR using ‘linearization primers’ (Sumo linear, Table 1). The sequence coding for the YadA O:9 N‐terminal peptide was amplified via PCR from the YadA O:9 ‐head domain construct (‘N‐terminal peptide insertion primers’, Table 1) [8]. Gibson cloning was used for insertion of the YadA O:9 N‐terminal peptide into the Sumo construct [49].…”

Section: Methodsmentioning

confidence: 99%

“…GAGs are part of the host ECM where they confer hydration [12,13]. The interaction between YadA O:9 and heparin is to date one of only two Type V secreted proteins that interact with a glycan [8,14]. Here, we elucidate the chemical and structural nature of the interaction between YadA O:9 and heparin.…”

Section: Introductionmentioning

confidence: 99%

A poly‐proline II helix in YadA from Yersinia enterocolitica serotype O:9 facilitates heparin binding through electrostatic interactions

Meuskens,

Kristiansen,

Bardiaux

et al. 2023

The FEBS Journal

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Poly‐proline II helices are secondary structure motifs frequently found in ligand binding sites. They exhibit increased flexibility and solvent exposure compared to the strongly hydrogen‐bonded α‐helices or β‐strands and can therefore easily be misinterpreted as completely unstructured regions with an extremely high rotational freedom. Here, we show that the adhesin YadA of Yersinia enterocolitica serotype O:9 contains a poly‐proline II helix interaction motif in the N‐terminal region. The motif is involved in the interaction of YadAO:9 with heparin, a host glycosaminoglycan. We show that the basic residues within the N‐terminal motif of YadA are required for electrostatic interactions with the sulphate groups of heparin. Biophysical methods including CD spectroscopy, solution‐state NMR, and SAXS all independently support the presence of a poly‐proline helix allowing YadAO:9 binding to the rigid heparin. Lastly, we show that host cells deficient in sulphation of heparin and heparan sulphate are not targeted by YadAO:9‐mediated adhesion. We speculate that the YadAO:9‐heparin interaction plays an important and highly strain‐specific role in the pathogenicity of Yersinia enterocolitica serotype O:9.

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“…Certain bacteria, such as Y. enterocolitica , assemble protein complexes called adhesins on their surface, which recognize specific molecular receptors of the host during colonization and are suitable markers for detection of the bacteria [ 32 , 33 ]. YadA from Y. enterocolitica is such an adhesin and is classified as a homotrimeric autotransporter [ 34 ], a class of adhesins that have been shown to enable many Gram-negative pathogens to interact with host extracellular matrix proteins, such as collagen, vitronectin, and fibronectin [ 35 , 36 ]. During an infection, YadA mediates the binding to epithelial and polymorphonuclear cells, and is also essential for autoagglutination [ 32 ].…”

Section: Introductionmentioning

confidence: 99%

Electrochemical Aptasensor for the Detection of the Key Virulence Factor YadA of Yersinia enterocolitica

et al. 2022

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New point-of-care (POC) diagnosis of bacterial infections are imperative to overcome the deficiencies of conventional methods, such as culture and molecular methods. In this study, we identified new aptamers that bind to the virulence factor Yersinia adhesin A (YadA) of Yersinia enterocolitica using cell-systematic evolution of ligands by exponential enrichment (cell-SELEX). Escherichia coli expressing YadA on the cell surface was used as a target cell. After eight cycles of selection, the final aptamer pool was sequenced by high throughput sequencing using the Illumina Novaseq platform. The sequencing data, analyzed using the Geneious software, was aligned, filtered and demultiplexed to obtain the key nucleotides possibly involved in the target binding. The most promising aptamer candidate, Apt1, bound specifically to YadA with a dissociation constant (Kd) of 11 nM. Apt1 was used to develop a simple electrochemical biosensor with a two-step, label-free design towards the detection of YadA. The sensor surface modifications and its ability to bind successfully and stably to YadA were confirmed by cyclic voltammetry, impedance spectroscopy and square wave voltammetry. The biosensor enabled the detection of YadA in a linear range between 7.0 × 104 and 7.0 × 107 CFU mL−1 and showed a square correlation coefficient >0.99. The standard deviation and the limit of detection was ~2.5% and 7.0 × 104 CFU mL−1, respectively. Overall, the results suggest that this novel biosensor incorporating Apt1 can potentially be used as a sensitive POC detection system to aid the diagnosis of Y. enterocolitica infections. Furthermore, this simple yet innovative approach could be replicated to select aptamers for other (bacterial) targets and to develop the corresponding biosensors for their detection.

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The Trimeric Autotransporter Adhesin YadA of Yersinia enterocolitica Serotype O:9 Binds Glycan Moieties

Cited by 9 publications

References 48 publications

Cell-density regulated adhesins contribute to early disease development and adhesion inRalstonia solanacearum

Cell-density regulated adhesins contribute to early disease development and adhesion inRalstonia solanacearum

A poly‐proline II helix in YadA from Yersinia enterocolitica serotype O:9 facilitates heparin binding through electrostatic interactions

Electrochemical Aptasensor for the Detection of the Key Virulence Factor YadA of Yersinia enterocolitica

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