The non-conserved region of MRP is involved in the virulence of<i>Streptococcus suis</i>serotype 2

Quan, Li Na; Fu, Yang; Ma, Caifeng; He, Yan; Yu, Yun William; Du, Dezhi; Yao, Huochun; Chen, Lu; Zhang, Wei

doi:10.1080/21505594.2017.1313373

Cited by 29 publications

(23 citation statements)

References 51 publications

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“…Interestingly, a new genotype was found among Canadian strains, named NA3. Since a role in virulence of the MRP protein has been recently highlighted [ 50 ], studies using more isolates from diseased pigs and ill patients will be required to evaluate the real significance of different genotypes of mrp and the correlation with the expression of the protein. None of the strains were epf + , which is in agreement with most previous serotype 9 strain studies [ 37 , 49 ].…”

Section: Discussionmentioning

confidence: 99%

Genomic comparisons of Streptococcus suis serotype 9 strains recovered from diseased pigs in Spain and Canada

Zheng¹,

Du²,

Qiu³

et al. 2018

Vet Res

101

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Streptococcus suis is one of the most important bacterial pathogens in the porcine industry and also a zoonotic agent. Serotype 9 is becoming one of the most prevalent serotypes within the S. suis population in certain European countries. In the present study, serotype 9 strains isolated from a country where infection due to this serotype is endemic (Spain), were compared to those recovered from Canada, where this serotype is rarely isolated from diseased pigs. For comparison purposes, strains from Brazil and the only strain isolated from a human case, in Thailand, were also incorporated. Firstly, sequence types (STs) were obtained followed by detection of putative virulence factors. Phylogenetic trees were constructed using the non-recombinant single nucleotide polymorphisms from core genomes of tested strains. Most Spanish strains were either ST123 or ST125, whereas Canadian strains were highly heterogeneous. However, the distribution of putative virulence factors was similar in both groups of strains. The fact that ST16 strains harbored more putative virulence genes and shared greater similarity with the genome of human serotype 2 strains suggests that they present a higher zoonotic and virulence potential than those from Canada and Spain. More than 80% of the strains included in this study carried genes associated with resistance to tetracycline, lincosamides and macrolides. Serotype 9 strains may be nearly 400 years old and have evolved in parallel into 2 lineages. The rapid population expansion of dominant lineage 1 occurred within the last 40 years probably due to the rapid development of the porcine industry.Electronic supplementary materialThe online version of this article (10.1186/s13567-017-0498-2) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Genomic comparisons of Streptococcus suis serotype 9 strains recovered from diseased pigs in Spain and Canada

Zheng¹,

Du²,

Qiu³

et al. 2018

Vet Res

101

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“…To determine the STEC components that Interact with EF-Tu, we examined the fibronectin-binding activities of rEF-Tu with far-WB analysis. The corresponding bands were observed in both reactions of rEF-Tu to anti-EF-Tu antibody (positive control) and to fibronectin, while no specific reaction was observed in the negative control MRP-D2, a non-related recombinant protein which was verified not to bind to fibronectin in a previous study (Li et al, 2017 ). The analysis indicated that rEF-Tu could specifically bind to fibronectin (Figure 7A ).…”

Section: Resultsmentioning

confidence: 82%

“…The cells were cultured in 24-well cell plates and fixed with cold methanol at −20°C for 20 min. Fixed cells were incubated with 100 μg of purified rEF-Tu or non-related recombinant protein HP07325, which was previously verified not to be involved in adhesion to epithelial cells (Li et al, 2017 ) at 37°C for 1 h. After three washes with PBS, the cells were incubated with His-tagged monoclonal antibody (Boster) at a 1:1,000 dilution. After washing three times with PBS, the cells were incubated with tetraethyl rhodamine isothiocyanate (TRITC)-tagged anti-IgG (Proteintech, 1:500 dilution) at 37°C for 30 min.…”

Section: Methodsmentioning

confidence: 99%

“…To explore if M. hyopneumoniae EF-Tu could bind to fibronectin, the protein-protein interactions method far-Western blot (far-WB) was performed. Total 20 μg recombinant proteins, including rEF-Tu and a negative control MRP-D2, which was previously verified not to bind to fibronectin (Li et al, 2017 ) were separated by SDS-PAGE and transferred to a polyvinylidene fluoride (PVDF) membrane (Li et al, 2015 ). After blocked with 5% (w/v) skimmed milk, the membrane was incubated with 5 μg/mL fibroenctin (Sigma), followed by incubation with rabbit anti-fibronectin antibody (Boster; 1 μg/mL) as the primary antibody, and horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (Boster; 1:5,000 dilution) as the secondary antibody.…”

Section: Methodsmentioning

confidence: 99%

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Elongation Factor Thermo Unstable (EF-Tu) Moonlights as an Adhesin on the Surface of Mycoplasma hyopneumoniae by Binding to Fibronectin

Yu¹,

Wang

Wang³

et al. 2018

Front. Microbiol.

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Mycoplasma hyopneumoniae is a colonizing respiratory pathogen that can cause great economic losses to the pig industry worldwide. Although putative virulence factors have been reported, the pathogenesis of this species remains unclear. Here, we used the virulent M. hyopneumoniae strain 168 to infect swine tracheal epithelial cells (STEC) to identify the infection-associated factors by two-dimensional electrophoresis (2-DE). Whole proteins of M. hyopneumoniae were obtained and compared with samples cultured in broth. Six differentially expressed proteins with an increase in abundance of ≥1.5 in the cell infection group were successfully identified. A String network of virulence-associated proteins showed that all the six differential abundance proteins were involved in virulence of M. hyopneumoniae. One of the most important upregulated hubs in this network, elongation factor thermo unstable (EF-Tu), which showed a relatively higher expression in M. hyopneumoniae-infected STEC and obtained a higher score on mass spectrometry was successfully recombined. In addition to its canonical enzymatic activities in protein synthesis, EF-Tu was also reported to be located on the cell surface as an important adhesin in many other pathogens. The cell surface location of EF-Tu was then observed in M. hyopneumoniae with flow cytometry. Recombinant EF-Tu (rEF-Tu) was found to be able to adhere to STEC and anti-rEF-Tu antibody enclosed M. hyopneumoniae decreased adherence to STEC. In addition, surface plasmon resonance (SPR) analysis showed that rEF-Tu could bind to fibronectin with a specific and moderately strong interaction, a dissociation constant (KD) of 605 nM. Furthermore, the block of fibronectin in STEC also decreased the binding of M. hyopneumoniae to the cell surface. Collectively, these data imply EF-Tu as an important adhesin of M. hyopneumoniae and fibronectin as an indispensable receptor on STEC. The binding between EF-Tu with fibronectin contributes to the adhesion of M. hyopneumoniae to STEC.HIGHLIGHTS Elongation factor thermo unstable (EF-Tu) exists on the cell surface of M. hyopneumoniae.EF-Tu moonlights as an adhesin of M. hyopneumoniae.The adhesive effect of EF-Tu is partly meditated by fibronectin.

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“…However, no ideal therapeutics or vaccine against S. suis infections is available thus far, although some researches showed homologous protection (Baums et al, 2009;Feng et al, 2014;Fittipaldi et al, 2012). To date, a multitude of virulence factors have been reported, such as MRP (Li et al, 2017b;Wisselink et al, 2001), SLY (Du et al, 2013;Jacobs et al, 1996), EF (Wisselink et al, 2001), enolase (Esgleas et al, 2009;Feng et al, 2009;Zhang et al, 2009), HtpsC (Li et al, 2015a), and HP0272 (Chen et al, 2010;Pian et al, 2012), but the pathogenesis of S. suis infections remains poorly understood (Fittipaldi et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Identification of novel pig and human immunoglobulin G-binding proteins and characterization of the binding regions of enolase from Streptococcus suis serotype 2

Quan

Guo

et al. 2020

Preprint

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Streptococcus suis, a major emerging pathogen in swine and humans, expresses immunoglobulin G (IgG)-binding proteins (IBPs), which contribute to the ability of organism to evasion of host immune system. The objective of this study was to identify novel pig IgG (pIgG) and human IgG (hIgG)-binding proteins and characterize the binding regions of enolase from Streptococcus suis serotype 2 (S. suis 2). Here, four pIgG-binding proteins (pIBPs) and five hIgG-binding proteins (hIBPs) were identified from S. suis 2 surface proteins by 2D-Far-western blot assays. All the newly captured proteins were expressed and further confirmed their binding activity to pIgG or hIgG by Far-western blot and dot blot. In addition to previously identified factor H, fibronectin, collagen, fibrinogen, plasminogen and laminin, we also found that both pIgG and hIgG can specifically interact with enolase. Binding assays indicated that interactions of S. suis 2 enolase with pIgG and hIgG is primarily mediated by the enolase C-terminal portion (Enolase-C, a.a. 142-432). We found that hIgG exhibited stronger binding ability to Enolase-C than pIgG. Further analysis of the C-terminal regions of enolase (Enolase-C1 and Enolase-C2) suggested that the C-terminus possessed two different binding domains with distinct host IgG proteins. Strikingly, we confirmed that pIgG interacted with the Enolase-C1 (a.a. 142-271) and hIgG interacted with the Enolase-C2 (a.a. 271-432). These observations of enolase provide interesting insights in the pathogenesis of S. suis infection.

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The non-conserved region of MRP is involved in the virulence ofStreptococcus suisserotype 2

Cited by 29 publications

References 51 publications

Genomic comparisons of Streptococcus suis serotype 9 strains recovered from diseased pigs in Spain and Canada

Genomic comparisons of Streptococcus suis serotype 9 strains recovered from diseased pigs in Spain and Canada

Elongation Factor Thermo Unstable (EF-Tu) Moonlights as an Adhesin on the Surface of Mycoplasma hyopneumoniae by Binding to Fibronectin

Identification of novel pig and human immunoglobulin G-binding proteins and characterization of the binding regions of enolase from Streptococcus suis serotype 2

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