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Enhancement of streptolysin O activity and intrinsic cytotoxic effects of the group A streptococcal toxin, NAD-glycohydrolase

Michos, Athanasios; Gryllos, Ioannis; Håkansson, Anders P LU ; Srivastava, Amit; Kokkotou, Efi and Wessels, Michael R (2006) In Journal of Biological Chemistry 281(12). p.8216-8223
Abstract

Streptolysin O (SLO) is a cholesterol-dependent cytolysin produced by the important human pathogen, group A Streptococcus (Streptococcus pyogenes or GAS). In addition to its cytolytic activity, SLO mediates the translocation of GAS NAD-glycohydrolase (NADase) into human epithelial cells in vitro. Production of both NADase and SLO is associated with augmented host cell injury beyond that produced by SLO alone, but the mechanism of enhanced cytotoxicity is not known. We have now shown that expression of NADase together with SLO dramatically enhanced the lytic activity of GAS culture supernatants for erythrocytes but had no effect on SLO-mediated poration of synthetic cholesterol-rich liposomes. This result revealed a previously unknown... (More)

Streptolysin O (SLO) is a cholesterol-dependent cytolysin produced by the important human pathogen, group A Streptococcus (Streptococcus pyogenes or GAS). In addition to its cytolytic activity, SLO mediates the translocation of GAS NAD-glycohydrolase (NADase) into human epithelial cells in vitro. Production of both NADase and SLO is associated with augmented host cell injury beyond that produced by SLO alone, but the mechanism of enhanced cytotoxicity is not known. We have now shown that expression of NADase together with SLO dramatically enhanced the lytic activity of GAS culture supernatants for erythrocytes but had no effect on SLO-mediated poration of synthetic cholesterol-rich liposomes. This result revealed a previously unknown contribution of NADase to the cytolytic activity associated with GAS production of SLO. Purified recombinant SLO bound NADase in vitro, supporting a specific, physical interaction of the two proteins. Exposure of human keratinocytes to wild-type GAS, but not to a NADase-deficient mutant strain, resulted in profound depletion of cellular NAD+ and ATP. Furthermore, expression of recombinant GAS NADase in yeast, in the absence of SLO, induced growth arrest, depletion of NAD+ and ATP, and cell death. These findings have provided evidence that the augmentation of SLO-mediated cytotoxicity by NADase is a consequence of depletion of host cell energy stores through the enzymatic action of NADase. Together, the results have provided mechanistic insight into the cytotoxic effects of a unique bipartite bacterial toxin.

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published
keywords
Adenosine Triphosphate, Bacterial Proteins, Bacterial Toxins, Blotting, Western, Cell Proliferation, Cells, Cultured, Chromatography, Affinity, DNA Primers, Dose-Response Relationship, Drug, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Escherichia coli, Exotoxins, Hemolysis, Humans, Immunoglobulin G, Keratinocytes, Liposomes, Microscopy, Fluorescence, NAD, NAD+ Nucleosidase, Octoxynol, Protein Binding, Protein Structure, Tertiary, Protein Transport, Recombinant Proteins, Reverse Transcriptase Polymerase Chain Reaction, Saccharomyces cerevisiae, Streptococcal Infections, Streptococcus pyogenes, Streptolysins, Time Factors, Virulence
in
Journal of Biological Chemistry
volume
281
issue
12
pages
8 pages
publisher
American Society for Biochemistry and Molecular Biology
external identifiers
  • Scopus:33646377649
ISSN
0021-9258
DOI
10.1074/jbc.M511674200
language
English
LU publication?
no
id
b58426be-1429-4abd-ac4f-4aae2064b030
date added to LUP
2016-05-21 13:52:07
date last changed
2016-11-01 08:40:17
@misc{b58426be-1429-4abd-ac4f-4aae2064b030,
  abstract     = {<p>Streptolysin O (SLO) is a cholesterol-dependent cytolysin produced by the important human pathogen, group A Streptococcus (Streptococcus pyogenes or GAS). In addition to its cytolytic activity, SLO mediates the translocation of GAS NAD-glycohydrolase (NADase) into human epithelial cells in vitro. Production of both NADase and SLO is associated with augmented host cell injury beyond that produced by SLO alone, but the mechanism of enhanced cytotoxicity is not known. We have now shown that expression of NADase together with SLO dramatically enhanced the lytic activity of GAS culture supernatants for erythrocytes but had no effect on SLO-mediated poration of synthetic cholesterol-rich liposomes. This result revealed a previously unknown contribution of NADase to the cytolytic activity associated with GAS production of SLO. Purified recombinant SLO bound NADase in vitro, supporting a specific, physical interaction of the two proteins. Exposure of human keratinocytes to wild-type GAS, but not to a NADase-deficient mutant strain, resulted in profound depletion of cellular NAD+ and ATP. Furthermore, expression of recombinant GAS NADase in yeast, in the absence of SLO, induced growth arrest, depletion of NAD+ and ATP, and cell death. These findings have provided evidence that the augmentation of SLO-mediated cytotoxicity by NADase is a consequence of depletion of host cell energy stores through the enzymatic action of NADase. Together, the results have provided mechanistic insight into the cytotoxic effects of a unique bipartite bacterial toxin.</p>},
  author       = {Michos, Athanasios and Gryllos, Ioannis and Håkansson, Anders P and Srivastava, Amit and Kokkotou, Efi and Wessels, Michael R},
  issn         = {0021-9258},
  keyword      = {Adenosine Triphosphate,Bacterial Proteins,Bacterial Toxins,Blotting, Western,Cell Proliferation,Cells, Cultured,Chromatography, Affinity,DNA Primers,Dose-Response Relationship, Drug,Electrophoresis, Polyacrylamide Gel,Enzyme-Linked Immunosorbent Assay,Escherichia coli,Exotoxins,Hemolysis,Humans,Immunoglobulin G,Keratinocytes,Liposomes,Microscopy, Fluorescence,NAD,NAD+ Nucleosidase,Octoxynol,Protein Binding,Protein Structure, Tertiary,Protein Transport,Recombinant Proteins,Reverse Transcriptase Polymerase Chain Reaction,Saccharomyces cerevisiae,Streptococcal Infections,Streptococcus pyogenes,Streptolysins,Time Factors,Virulence},
  language     = {eng},
  month        = {03},
  number       = {12},
  pages        = {8216--8223},
  publisher    = {ARRAY(0x7e25eb8)},
  series       = {Journal of Biological Chemistry},
  title        = {Enhancement of streptolysin O activity and intrinsic cytotoxic effects of the group A streptococcal toxin, NAD-glycohydrolase},
  url          = {http://dx.doi.org/10.1074/jbc.M511674200},
  volume       = {281},
  year         = {2006},
}