The Extracellular Domain of the β<sub>2</sub> Integrin β Subunit (CD18) Is Sufficient for <named-content content-type="genus-species">Escherichia coli</named-content> Hemolysin and <named-content content-type="genus-species">Aggregatibacter actinomycetemcomitans</named-content> Leukotoxin Cytotoxic Activity

ABSTRACT The Escherichia coli hemolysin (HlyA) is a pore-forming exotoxin associated with severe complications of human urinary tract infections. HlyA is the prototype of the repeats-in-toxin (RTX) family, which includes LtxA from Aggregatibacter actinomycetemcomitans, a periodontal pathogen. The ex...

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Autores principales:	Laura C. Ristow, Vy Tran, Kevin J. Schwartz, Lillie Pankratz, Andrew Mehle, John-Demian Sauer, Rodney A. Welch
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	cytotoxicity hemolysin integrins leukotoxin RTX toxin Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/07258d37f51446f99ca294545c1ad19d
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spelling	oai:doaj.org-article:07258d37f51446f99ca294545c1ad19d2021-11-15T16:22:09ZThe Extracellular Domain of the β<sub>2</sub> Integrin β Subunit (CD18) Is Sufficient for <named-content content-type="genus-species">Escherichia coli</named-content> Hemolysin and <named-content content-type="genus-species">Aggregatibacter actinomycetemcomitans</named-content> Leukotoxin Cytotoxic Activity10.1128/mBio.01459-192150-7511https://doaj.org/article/07258d37f51446f99ca294545c1ad19d2019-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01459-19https://doaj.org/toc/2150-7511ABSTRACT The Escherichia coli hemolysin (HlyA) is a pore-forming exotoxin associated with severe complications of human urinary tract infections. HlyA is the prototype of the repeats-in-toxin (RTX) family, which includes LtxA from Aggregatibacter actinomycetemcomitans, a periodontal pathogen. The existence and requirement for a host cell receptor for these toxins are controversial. We performed an unbiased forward genetic selection in a mutant library of human monocytic cells, U-937, for host factors involved in HlyA cytotoxicity. The top candidate was the β2 integrin β subunit. Δβ2 cell lines are approximately 100-fold more resistant than wild-type U-937 cells to HlyA, but remain sensitive to HlyA at high concentrations. Similarly, Δβ2 cells are more resistant than wild-type U-937 cells to LtxA, as Δβ2 cells remain LtxA resistant even at >1,000-fold-higher concentrations of the toxin. Loss of any single β2 integrin α subunit, or even all four α subunits together, does not confer resistance to HlyA. HlyA and LtxA bind to the β2 subunit, but not to αL, αM, or αX in far-Western blots. Genetic complementation of Δβ2 cells with either β2 or β2 with a cytoplasmic tail deletion restores HlyA and LtxA sensitivity, suggesting that β2 integrin signaling is not required for cytotoxicity. Finally, β2 mutations do not alter sensitivity to unrelated pore-forming toxins, as wild-type or Δβ2 cells are equally sensitive to Staphylococcus aureus α-toxin and Proteus mirabilis HpmA. Our studies show two RTX toxins use the β2 integrin β subunit alone to facilitate cytotoxicity, but downstream integrin signaling is dispensable. IMPORTANCE Urinary tract infections are one of the most common bacterial infections worldwide. Uropathogenic Escherichia coli strains are responsible for more than 80% of community-acquired urinary tract infections. Although we have known for nearly a century that severe infections stemming from urinary tract infections, including kidney or bloodstream infections are associated with expression of a toxin, hemolysin, from uropathogenic Escherichia coli, how hemolysin functions to enhance virulence is unknown. Our research defines the interaction of hemolysin with the β2 integrin, a human white cell adhesion molecule, as a potential therapeutic target during urinary tract infections. The E. coli hemolysin is the prototype for a toxin family (RTX family) produced by a wide array of human and animal pathogens. Our work extends to the identification and characterization of the receptor for an additional member of the RTX family, suggesting that this interaction may be broadly conserved throughout the RTX toxin family.Laura C. RistowVy TranKevin J. SchwartzLillie PankratzAndrew MehleJohn-Demian SauerRodney A. WelchAmerican Society for MicrobiologyarticlecytotoxicityhemolysinintegrinsleukotoxinRTX toxinMicrobiologyQR1-502ENmBio, Vol 10, Iss 4 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	cytotoxicity hemolysin integrins leukotoxin RTX toxin Microbiology QR1-502
spellingShingle	cytotoxicity hemolysin integrins leukotoxin RTX toxin Microbiology QR1-502 Laura C. Ristow Vy Tran Kevin J. Schwartz Lillie Pankratz Andrew Mehle John-Demian Sauer Rodney A. Welch The Extracellular Domain of the β<sub>2</sub> Integrin β Subunit (CD18) Is Sufficient for <named-content content-type="genus-species">Escherichia coli</named-content> Hemolysin and <named-content content-type="genus-species">Aggregatibacter actinomycetemcomitans</named-content> Leukotoxin Cytotoxic Activity
description	ABSTRACT The Escherichia coli hemolysin (HlyA) is a pore-forming exotoxin associated with severe complications of human urinary tract infections. HlyA is the prototype of the repeats-in-toxin (RTX) family, which includes LtxA from Aggregatibacter actinomycetemcomitans, a periodontal pathogen. The existence and requirement for a host cell receptor for these toxins are controversial. We performed an unbiased forward genetic selection in a mutant library of human monocytic cells, U-937, for host factors involved in HlyA cytotoxicity. The top candidate was the β2 integrin β subunit. Δβ2 cell lines are approximately 100-fold more resistant than wild-type U-937 cells to HlyA, but remain sensitive to HlyA at high concentrations. Similarly, Δβ2 cells are more resistant than wild-type U-937 cells to LtxA, as Δβ2 cells remain LtxA resistant even at >1,000-fold-higher concentrations of the toxin. Loss of any single β2 integrin α subunit, or even all four α subunits together, does not confer resistance to HlyA. HlyA and LtxA bind to the β2 subunit, but not to αL, αM, or αX in far-Western blots. Genetic complementation of Δβ2 cells with either β2 or β2 with a cytoplasmic tail deletion restores HlyA and LtxA sensitivity, suggesting that β2 integrin signaling is not required for cytotoxicity. Finally, β2 mutations do not alter sensitivity to unrelated pore-forming toxins, as wild-type or Δβ2 cells are equally sensitive to Staphylococcus aureus α-toxin and Proteus mirabilis HpmA. Our studies show two RTX toxins use the β2 integrin β subunit alone to facilitate cytotoxicity, but downstream integrin signaling is dispensable. IMPORTANCE Urinary tract infections are one of the most common bacterial infections worldwide. Uropathogenic Escherichia coli strains are responsible for more than 80% of community-acquired urinary tract infections. Although we have known for nearly a century that severe infections stemming from urinary tract infections, including kidney or bloodstream infections are associated with expression of a toxin, hemolysin, from uropathogenic Escherichia coli, how hemolysin functions to enhance virulence is unknown. Our research defines the interaction of hemolysin with the β2 integrin, a human white cell adhesion molecule, as a potential therapeutic target during urinary tract infections. The E. coli hemolysin is the prototype for a toxin family (RTX family) produced by a wide array of human and animal pathogens. Our work extends to the identification and characterization of the receptor for an additional member of the RTX family, suggesting that this interaction may be broadly conserved throughout the RTX toxin family.
format	article
author	Laura C. Ristow Vy Tran Kevin J. Schwartz Lillie Pankratz Andrew Mehle John-Demian Sauer Rodney A. Welch
author_facet	Laura C. Ristow Vy Tran Kevin J. Schwartz Lillie Pankratz Andrew Mehle John-Demian Sauer Rodney A. Welch
author_sort	Laura C. Ristow
title	The Extracellular Domain of the β<sub>2</sub> Integrin β Subunit (CD18) Is Sufficient for <named-content content-type="genus-species">Escherichia coli</named-content> Hemolysin and <named-content content-type="genus-species">Aggregatibacter actinomycetemcomitans</named-content> Leukotoxin Cytotoxic Activity
title_short	The Extracellular Domain of the β<sub>2</sub> Integrin β Subunit (CD18) Is Sufficient for <named-content content-type="genus-species">Escherichia coli</named-content> Hemolysin and <named-content content-type="genus-species">Aggregatibacter actinomycetemcomitans</named-content> Leukotoxin Cytotoxic Activity
title_full	The Extracellular Domain of the β<sub>2</sub> Integrin β Subunit (CD18) Is Sufficient for <named-content content-type="genus-species">Escherichia coli</named-content> Hemolysin and <named-content content-type="genus-species">Aggregatibacter actinomycetemcomitans</named-content> Leukotoxin Cytotoxic Activity
title_fullStr	The Extracellular Domain of the β<sub>2</sub> Integrin β Subunit (CD18) Is Sufficient for <named-content content-type="genus-species">Escherichia coli</named-content> Hemolysin and <named-content content-type="genus-species">Aggregatibacter actinomycetemcomitans</named-content> Leukotoxin Cytotoxic Activity
title_full_unstemmed	The Extracellular Domain of the β<sub>2</sub> Integrin β Subunit (CD18) Is Sufficient for <named-content content-type="genus-species">Escherichia coli</named-content> Hemolysin and <named-content content-type="genus-species">Aggregatibacter actinomycetemcomitans</named-content> Leukotoxin Cytotoxic Activity
title_sort	extracellular domain of the β<sub>2</sub> integrin β subunit (cd18) is sufficient for <named-content content-type="genus-species">escherichia coli</named-content> hemolysin and <named-content content-type="genus-species">aggregatibacter actinomycetemcomitans</named-content> leukotoxin cytotoxic activity
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/07258d37f51446f99ca294545c1ad19d
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