Extracellular Nucleotide Catabolism by the Group B Streptococcus Ectonucleotidase NudP Increases Bacterial Survival in Blood

Firon, Arnaud; Dinis, Márcia; Raynal, Bertrand; Poyart, Claire; Trieu-Cuot, Patrick; Kaminski, P.A.

doi:10.1074/jbc.m113.545632

Cited by 37 publications

(45 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…zooepidemicus virulence decreased, and its ability to degrade NETs and generate deoxyadenosine was lost. Additionally, ENuc and 5Nuc possess the ability to hydrolyze AMP, ADP, and ATP to produce the immunomodulatory substrate adenosine, similar to Nt5e produced by Streptococcus sanguinis and AdsA produced by Staphylococcus aureus but unlike S5nA produced by Streptococcus pyogenes and NudP produced by Streptococcus agalactiae, which demonstrate limited specificity for AMP and ADP (21,24,25). Adenosine suppresses neutrophil activation by occupying the A2a receptor on neutrophils and promoting bacterial survival in the host bloodstream (26).…”

Section: Discussionmentioning

confidence: 99%

Extracellular Nucleases of Streptococcus equi subsp. zooepidemicus Degrade Neutrophil Extracellular Traps and Impair Macrophage Activity of the Host

Guo

Fan

2017

Appl Environ Microbiol

View full text Add to dashboard Cite

The pathogen Streptococcus equi subsp. zooepidemicus is associated with a wide range of animals, including humans, and outbreaks frequently occur in pigs, equines, and goats. Thus far, few studies have assessed interactions between the host immune system and S. equi subsp. zooepidemicus and how these interactions explain the wide host spectrum of S. equi subsp. zooepidemicus. Neutrophils, the first line of innate immunity, possess a defense mechanism called neutrophil extracellular traps (NETs), which primarily consist of DNA and granule proteins that trap bacteria via charge interactions. Extracellular nucleases play important roles in the degradation of the DNA backbone of NETs. Here, two related extracellular nucleases, nuclease and 5=-nucleotidase (named ENuc and 5Nuc, respectively, in this study), were identified as being encoded by the SESEC_RS04165 gene and the SESEC_RS05720 gene (named ENuc and 5Nuc, respectively), and three related gene deletion mutant strains, specifically, the single-mutant ⌬ENuc and ⌬5Nuc strains and the double-mutant ⌬ENuc ⌬5Nuc strain, were constructed. The ⌬ENuc and ⌬5Nuc single-mutant strains and the ⌬ENuc ⌬5Nuc double-mutant strain demonstrated lower virulence than wild-type S. equi subsp. zooepidemicus when the mouse survival rate was evaluated postinfection. Furthermore, wild-type S. equi subsp. zooepidemicus more frequently traversed the bloodstream and transferred to other organs. Wild-type S. equi subsp. zooepidemicus induced fewer NETs and was able to survive in NETs, whereas only 40% of the ⌬ENuc ⌬5Nuc double-mutant cells survived. S. equi subsp. zooepidemicus degraded the NET DNA backbone and produced deoxyadenosine, primarily through the action of ENuc and/or 5Nuc. However, the double-mutant ⌬ENuc ⌬5Nuc strain lost the ability to degrade NETs into deoxyadenosine. Deoxyadenosine decreased RAW 264.7 cell phagocytosis to 40% of that of normal macrophages.IMPORTANCE Streptococcus equi subsp. zooepidemicus causes serious bacteremia in its hosts. However, little is known about how S. equi subsp. zooepidemicus interacts with the host innate immune system, particularly innate cells found in the blood. S. equi subsp. zooepidemicus is capable of evading NET-mediated killing via the actions of its potent extracellular nucleases, ENuc and 5Nuc, which directly degrade the NET DNA backbone to deoxyadenosine. In previous studies, other pathogens have required the synergism of nuclease and 5=-nucleotidase to engage in this self-protective process; however, ENuc and 5Nuc both possess nuclease activity and 5=-nucleotidase activity, highlighting the novelty of this discovery. Furthermore, deoxyadenosine impairs phagocytosis but not the intracellular bactericidal activity of macrophages. Here we describe a novel mechanism for S. equi subsp. zooepidemicus extracellular nucleases in NET degradation, which may provide new insights into the pathogen immune evasion mechanism and the prevention and treatment of bacterial disease.

show abstract

Section: Discussionmentioning

confidence: 99%

Extracellular Nucleases of Streptococcus equi subsp. zooepidemicus Degrade Neutrophil Extracellular Traps and Impair Macrophage Activity of the Host

Guo

Fan

2017

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…Nucleotide biosynthesis pathways are critical for growth of bacteria (E. coli, Salmonella typhimurium, Bacillus anthracis) in human blood, and the enzymes involved are considered as targets for development of new antibiotics (Samant et al, 2008). Extracellular nucleotide metabolism is a well-known mediator of bacterial virulence (Firon et al, 2014). Over 50 % of FDA-approved antiviral and anticancer drugs are nucleoside or nucleobase analogues, and the potential use of these drugs as antibiotics is receiving increasing attention (Sun & Wang, 2013).…”

Section: Introductionmentioning

confidence: 99%

EF0176 and EF0177 from Enterococcus faecalis V583 are substrate-binding lipoproteins involved in ABC transporter mediated ribonucleoside uptake

et al. 2015

View full text Add to dashboard Cite

One of the ABC transporter systems in Enterococcus faecalis V583 is encoded by the ef0176-ef0180 gene cluster, which differs from orthologous operons in related bacteria in that it contains two genes putatively encoding substrate-binding proteins (SBPs). These SBPs, EF0176 and EF0177, have previously been identified on the surface of E. faecalis. By phenotypic studies of single and double knockout mutants, we show here that EF0176 and EF0177 are specific for ribonucleosides and, by inference, that the EF0176-EF0180 ABC transporter plays a role in nucleoside uptake. The specificity of the SBPs was mapped using growth experiments on a medium, RPMI 1640, that only supports growth of E. faecalis when supplemented with purine nucleosides or their corresponding bases. This analysis was complemented by studies with toxic fluorinated pyrimidine ribonucleoside analogues and competition experiments. The data show that EF0176 and EF0177 have broad and overlapping, but not identical, substrate specificities and that they, together, are likely to bind and facilitate the transport of all common ribonucleosides. Comparative sequence analysis and inspection of an available crystal structure of an orthologue, PnrA from Treponema pallidum, showed that the strongest binding interactions between the protein and the ligand involve the ribose moiety and that sequence variation in the binding site primarily affects interactions with the base. This explains both the broad substrate specificity of these binding proteins and the observed variations therein. The presence of two SBPs in this nucleoside ABC transporter system in E. faecalis may improve the bacterium's ability to scavenge nucleosides.

show abstract

“…Each of these enzymes was shown to convert AMP into the immunomodulator adenosine, facilitating the evasion of the bacteria from the host immune response (20 -23, 25, 26). In addition, the S. aureus AdsA and NudP from S. agalactiae are also able to convert dAMP into deoxyadenosine, which has cytotoxic effects on macrophages and monocytes (22,25). Here, we report the discovery of a novel nucleotidase from the human pathogen S. pyogenes, which we name S5nA.…”

Section: Discussionmentioning

confidence: 96%

“…Identification of S5nA-All previously reported bacterial nucleotidases produced by Gram-positive bacteria are cell wallanchored proteins with a characteristic C-terminal cell wall anchoring domain and a LPXTG sorting motif that is recognized by the housekeeping sortase A enzyme (21,23,25). Using the S. pyogenes SF370 (serotype M1) genome sequence (31), we searched for all proteins with typical Gram-positive cell wall anchor domains and found an open reading frame (spy0872) that was annotated as putative secreted 5Ј-nucleotidase (AAK33792).…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Streptococcal 5′-Nucleotidase A (S5nA), a Novel Streptococcus pyogenes Virulence Factor That Facilitates Immune Evasion

Zheng¹,

Khemlani²,

Lorenz

et al. 2015

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background: 5Ј-Nucleotidases are important virulence factors found in several bacterial pathogens. Results: Streptococcal 5Ј-nucleotidase A (S5nA) generated immunomodulatory molecules adenosine and deoxyadenosine and rescued Lactococcus lactis in a blood killing assay. Conclusion: S5nA is a novel Streptococcus pyogenes virulence factor that facilitates immune evasion from the host. Significance: S5nA might be a target for developing new therapeutics or vaccines.

show abstract

Extracellular Nucleotide Catabolism by the Group B Streptococcus Ectonucleotidase NudP Increases Bacterial Survival in Blood

Cited by 37 publications

References 53 publications

Extracellular Nucleases of Streptococcus equi subsp. zooepidemicus Degrade Neutrophil Extracellular Traps and Impair Macrophage Activity of the Host

Extracellular Nucleases of Streptococcus equi subsp. zooepidemicus Degrade Neutrophil Extracellular Traps and Impair Macrophage Activity of the Host

EF0176 and EF0177 from Enterococcus faecalis V583 are substrate-binding lipoproteins involved in ABC transporter mediated ribonucleoside uptake

Streptococcal 5′-Nucleotidase A (S5nA), a Novel Streptococcus pyogenes Virulence Factor That Facilitates Immune Evasion

Contact Info

Product

Resources

About