NAD<sup>+</sup>-glycohydrolase productivity of haemolytic streptococci assayed by a simple fluorescent method and its relation to T serotype

Karasawa, Tadahiro; Yamakawa, Kiyotaka; Tanaka, Daisuke; Gyobu, Yotaku; Nakamura, Shinichi

doi:10.1111/j.1574-6968.1995.tb07538.x

Cited by 15 publications

(15 citation statements)

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“…The expected phenotypes for 188NADase Ϫ were confirmed in assays for SLO-mediated hemolysis and NAD glycohydrolase (NADase) enzymatic activity (19,20).…”

Section: Methodsmentioning

confidence: 86%

Cytolysin-dependent evasion of lysosomal killing

Håkansson

Bentley

Shakhnovic

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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Local host defenses limit proliferation and systemic spread of pathogenic bacteria from sites of mucosal colonization. For pathogens such as streptococci that fail to grow intracellularly, internalization and killing by epithelial cells contribute to the control of bacterial growth and dissemination. Here, we show that group A Streptococcus (GAS), the agent of streptococcal sore throat and invasive soft tissue infections, evades internalization and intracellular killing by pharyngeal epithelial cells. Production of the cholesterol-binding cytotoxin streptolysin O (SLO) prevented internalization of GAS into lysosomes. In striking contrast, GAS rendered defective in production of SLO were internalized directly or rapidly transported into lysosomes, where they were killed by a pH-dependent mechanism. Because SLO is the prototype of cholesterol-dependent cytolysins produced by many Grampositive bacteria, cytolysin-mediated evasion of lysosomal killing may be a general mechanism to protect such pathogens from clearance by host epithelial cells.Streptococcus pyogenes ͉ streptolysin O ͉ virulence E pithelial cells of mucosal surfaces constitute an early line of defense against infecting microorganisms. These cells protect the host not only by acting as a physical barrier but also by secreting antimicrobial molecules and by recruiting professional immune cells to the site of infection (1-5). Also, epithelial cells have the capacity to internalize and kill bacteria. Extracellular pathogens such as streptococci, Staphylococcus aureus, and Pseudomonas aeruginosa can be internalized by epithelial cells but do not appear to multiply in the intracellular environment (6-11). Rather, internalized bacteria are killed over time. Therefore, to persist at the mucosal surface, extracellular bacterial pathogens must evade internalization and killing by epithelial cells.In this study, we investigated the effects of streptolysin O (SLO) on the interactions of group A Streptococcus (GAS), the agent of streptococcal sore throat and severe, invasive, or ''flesh-eating'' infections, with human oropharyngeal epithelial cells. SLO is the prototype of a family of cholesterol-binding cytotoxins produced by many pathogenic Gram-positive bacteria including Streptococcus pneumoniae (pneumolysin), Listeria monocytogenes (listeriolysin O), Clostridium perfringens (perfringolysin O), and Bacillus anthracis (anthrolysin) (12-14). The cholesterol-binding cytotoxins share the property of poreforming activity for a broad range of cholesterol-containing cell membranes (12, 15). However, it remains unknown whether they serve a common functional role in pathogen-host biology for the diverse species that produce them. Here, we report that SLO is a critical modulator of GAS internalization, intracellular trafficking, and bacterial killing by human oropharyngeal keratinocytes. We found that SLO prevented direct uptake of GAS into lysosomes and efficient pH-dependent intracellular killing. On the basis of these findings, we propose that SLO enhances GAS surviv...

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“…The expected phenotypes for 188NADase Ϫ were confirmed in assays for SLO-mediated hemolysis and NAD glycohydrolase (NADase) enzymatic activity (19,20).…”

Section: Methodsmentioning

confidence: 86%

Cytolysin-dependent evasion of lysosomal killing

Håkansson

Bentley

Shakhnovic

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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“…Clinical streptococcal isolates often secrete NADases (24), and an NADase activity distinct from cholera toxin has been described as an extracellular product of V. cholerae (52). Also, an ORF resembling HvnA and HvnB occurs in the P. aeruginosa genome, suggesting that this organism may also secrete an NADase.…”

Section: Fig 6 Product Analysis Of Nadmentioning

confidence: 99%

“…Also, an ORF resembling HvnA and HvnB occurs in the P. aeruginosa genome, suggesting that this organism may also secrete an NADase. Each of these bacterial species, as well as V. fischeri, colonizes the extracellular surface of animal cells, leading us and others (24) to speculate that secretion of NADases may play a role in host colonization or bacterium-animal signaling. NADases could function in animal-bacterium interactions by any of a number of mechanisms.…”

Section: Fig 6 Product Analysis Of Nadmentioning

confidence: 99%

“…Thus, the assay used to identify HvnA and HvnB did not distinguish between ARTase or NADase activity. Secreted bacterial ARTases clearly mediate defined effects on certain animal host cells (15,(39)(40)(41), but the role, if any, of secreted NADases in bacterium-host interactions is unknown beyond the observations that clinical streptococcal isolates and Vibrio cholerae secrete NADases (24,52).…”

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confidence: 99%

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Vibrio fischeriGeneshvnAandhvnBEncode Secreted NAD⁺-Glycohydrolases

2001

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HvnA and HvnB are proteins secreted by Vibrio fischeri ES114, an extracellular light organ symbiont of the squid Euprymna scolopes, that catalyze the transfer of ADP-ribose from NAD ؉ to polyarginine. Based on this activity, HvnA and HvnB were presumptively designated mono-ADP-ribosyltransferases (ARTases), and it was hypothesized that they mediate bacterium-host signaling. We have cloned hvnA and hvnB from strain ES114. hvnA appears to be expressed as part of a four-gene operon, whereas hvnB is monocistronic. The predicted HvnA and HvnB amino acid sequences are 46% identical to one another and share 44% and 34% identity,

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“…This secreted toxin has an enzymatic activity (NADase) that cleaves the glycosidic bond of ␤-NAD ϩ to produce nicotinamide and ADP-ribose. All S. pyogenes strains examined to date possess the gene that encodes SPN (spn), but some strains produce a SPN that lacks detectable NADase activity (1,30,36,42). Since there is evidence that SPNЈs robust NADase activity contributes to virulence (4,43,52,56), the existence of NADase-deficient SPN has yet to be explained.…”

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confidence: 99%

Variation inStreptococcus pyogenesNAD⁺Glycohydrolase Is Associated with Tissue Tropism

2010

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Streptococcus pyogenes is an important pathogen that causes a variety of diseases. The most common infections involve the throat (pharyngitis) or skin (impetigo); however, the factors that determine tissue tropism and severity are incompletely understood. The S. pyogenes NAD ؉ glycohydrolase (SPN) is a virulence factor that has been implicated in contributing to the pathogenesis of severe infections. However, the role of SPN in determining the bacterium's tissue tropism has not been evaluated. In this report, we examine the sequences of spn and its endogenous inhibitor ifs from a worldwide collection of S. pyogenes strains. Analysis of average pairwise nucleotide diversity, average number of nucleotide differences, and ratio of nonsynonymous to synonymous substitutions revealed significant diversity in spn and ifs. Application of established models of molecular evolution shows that SPN is evolving under positive selection and diverging into NAD ؉ glycohydrolase (NADase)-active and -inactive subtypes. Additionally, the NADase-inactive SPN subtypes maintain the characteristics of a functional gene while ifs becomes a pseudogene. Thus, NADase-inactive SPN continues to evolve under functional constraint. Furthermore, NADase activity did not correlate with invasive disease in our collection but was associated with tissue tropism. The ability to cause infection at both the pharynx and the skin ("generalist" strains) is correlated with NADase-active SPN, while the preference for causing infection at either the throat or the skin ("specialist" strains) is associated with NADase-inactive SPN. These findings suggest that SPN has a NADase-independent function and prompt a reevaluation of the role of SPN in streptococcal pathogenesis.

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NAD⁺-glycohydrolase productivity of haemolytic streptococci assayed by a simple fluorescent method and its relation to T serotype

Cited by 15 publications

References 10 publications

Cytolysin-dependent evasion of lysosomal killing

Cytolysin-dependent evasion of lysosomal killing

Vibrio fischeriGeneshvnAandhvnBEncode Secreted NAD⁺-Glycohydrolases

Variation inStreptococcus pyogenesNAD⁺Glycohydrolase Is Associated with Tissue Tropism

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NAD+-glycohydrolase productivity of haemolytic streptococci assayed by a simple fluorescent method and its relation to T serotype

Cited by 15 publications

References 10 publications

Cytolysin-dependent evasion of lysosomal killing

Cytolysin-dependent evasion of lysosomal killing

Vibrio fischeriGeneshvnAandhvnBEncode Secreted NAD+-Glycohydrolases

Variation inStreptococcus pyogenesNAD+Glycohydrolase Is Associated with Tissue Tropism

Contact Info

Product

Resources

About

NAD⁺-glycohydrolase productivity of haemolytic streptococci assayed by a simple fluorescent method and its relation to T serotype

Vibrio fischeriGeneshvnAandhvnBEncode Secreted NAD⁺-Glycohydrolases

Variation inStreptococcus pyogenesNAD⁺Glycohydrolase Is Associated with Tissue Tropism