Cellular Activity of <i>Salmonella</i> Typhimurium ArtAB Toxin and Its Receptor-Binding Subunit
Salmonellosis is among the most reported foodborne illnesses in the United States. The <i>Salmonella</i><i>enterica</i> Typhimurium DT104 phage type, which is associated with multidrug-resistant disease in humans and animals, possesses an ADP-ribosylating toxin called ArtAB....
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MDPI AG
2021-08-01
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author | Elise Overgaard Brad Morris Omid Mohammad Mousa Emily Price Adriana Rodriguez Leyla Cufurovic Richard S. Beard Juliette K. Tinker |
author_facet | Elise Overgaard Brad Morris Omid Mohammad Mousa Emily Price Adriana Rodriguez Leyla Cufurovic Richard S. Beard Juliette K. Tinker |
author_sort | Elise Overgaard |
collection | DOAJ |
description | Salmonellosis is among the most reported foodborne illnesses in the United States. The <i>Salmonella</i><i>enterica</i> Typhimurium DT104 phage type, which is associated with multidrug-resistant disease in humans and animals, possesses an ADP-ribosylating toxin called ArtAB. Full-length <i>art</i>AB has been found on a number of broad-host-range non-typhoidal <i>Salmonella</i> species and serovars. ArtAB is also homologous to many AB<sub>5</sub> toxins from diverse Gram-negative pathogens, including cholera toxin (CT) and pertussis toxin (PT), and may be involved in <i>Salmonella</i> pathogenesis, however, in vitro cellular toxicity of ArtAB has not been characterized. <i>art</i>AB was cloned into <i>E. coli</i> and initially isolated using a histidine tag (ArtABHIS) and nickel chromatography. ArtABHIS was found to bind to African green monkey kidney epithelial (Vero) cells using confocal microscopy and to interact with glycans present on fetuin and monosialotetrahexosylganglioside (GM1) using ELISA. Untagged, or native, holotoxin (ArtAB), and the pentameric receptor-binding subunit (ArtB) were purified from <i>E. coli</i> using fetuin and <span style="font-variant: small-caps;">d</span>-galactose affinity chromatography. ArtAB and ArtB metabolic and cytotoxic activities were determined using Vero and Chinese hamster ovary (CHO) epithelial cells. Vero cells were more sensitive to ArtAB, however, incubation with both cell types revealed only partial cytotoxicity over 72 h, similar to that induced by CT. ArtAB induced a distinctive clustering phenotype on CHO cells over 72 h, similar to PT, and an elongated phenotype on Vero cells, similar to CT. The ArtB binding subunit alone also had a cytotoxic effect on CHO cells and induced morphological rounding. Results indicate that this toxin induces distinctive cellular outcomes. Continued biological characterization of ArtAB will advance efforts to prevent disease caused by non-typhoidal <i>Salmonella</i>. |
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spelling | doaj.art-213781e16a0e43ce93b9cdb56754d21d2023-11-22T15:30:45ZengMDPI AGToxins2072-66512021-08-0113959910.3390/toxins13090599Cellular Activity of <i>Salmonella</i> Typhimurium ArtAB Toxin and Its Receptor-Binding SubunitElise Overgaard0Brad Morris1Omid Mohammad Mousa2Emily Price3Adriana Rodriguez4Leyla Cufurovic5Richard S. Beard6Juliette K. Tinker7Biomolecular Sciences Graduate Program, Boise State University, Boise, ID 83725, USADepartment of Biology, Boise State University, Boise, ID 83725, USADepartment of Biology, Boise State University, Boise, ID 83725, USAIdaho Veterans Research and Education Foundation, Infectious Diseases Section, Boise, ID 83702, USADepartment of Biology, Boise State University, Boise, ID 83725, USADepartment of Biology, Boise State University, Boise, ID 83725, USABiomolecular Research Center, Boise State University, Boise, ID 83725, USABiomolecular Sciences Graduate Program, Boise State University, Boise, ID 83725, USASalmonellosis is among the most reported foodborne illnesses in the United States. The <i>Salmonella</i><i>enterica</i> Typhimurium DT104 phage type, which is associated with multidrug-resistant disease in humans and animals, possesses an ADP-ribosylating toxin called ArtAB. Full-length <i>art</i>AB has been found on a number of broad-host-range non-typhoidal <i>Salmonella</i> species and serovars. ArtAB is also homologous to many AB<sub>5</sub> toxins from diverse Gram-negative pathogens, including cholera toxin (CT) and pertussis toxin (PT), and may be involved in <i>Salmonella</i> pathogenesis, however, in vitro cellular toxicity of ArtAB has not been characterized. <i>art</i>AB was cloned into <i>E. coli</i> and initially isolated using a histidine tag (ArtABHIS) and nickel chromatography. ArtABHIS was found to bind to African green monkey kidney epithelial (Vero) cells using confocal microscopy and to interact with glycans present on fetuin and monosialotetrahexosylganglioside (GM1) using ELISA. Untagged, or native, holotoxin (ArtAB), and the pentameric receptor-binding subunit (ArtB) were purified from <i>E. coli</i> using fetuin and <span style="font-variant: small-caps;">d</span>-galactose affinity chromatography. ArtAB and ArtB metabolic and cytotoxic activities were determined using Vero and Chinese hamster ovary (CHO) epithelial cells. Vero cells were more sensitive to ArtAB, however, incubation with both cell types revealed only partial cytotoxicity over 72 h, similar to that induced by CT. ArtAB induced a distinctive clustering phenotype on CHO cells over 72 h, similar to PT, and an elongated phenotype on Vero cells, similar to CT. The ArtB binding subunit alone also had a cytotoxic effect on CHO cells and induced morphological rounding. Results indicate that this toxin induces distinctive cellular outcomes. Continued biological characterization of ArtAB will advance efforts to prevent disease caused by non-typhoidal <i>Salmonella</i>.https://www.mdpi.com/2072-6651/13/9/599<i>Salmonella</i> TyphimuriumAB<sub>5</sub> toxinArtAB |
spellingShingle | Elise Overgaard Brad Morris Omid Mohammad Mousa Emily Price Adriana Rodriguez Leyla Cufurovic Richard S. Beard Juliette K. Tinker Cellular Activity of <i>Salmonella</i> Typhimurium ArtAB Toxin and Its Receptor-Binding Subunit Toxins <i>Salmonella</i> Typhimurium AB<sub>5</sub> toxin ArtAB |
title | Cellular Activity of <i>Salmonella</i> Typhimurium ArtAB Toxin and Its Receptor-Binding Subunit |
title_full | Cellular Activity of <i>Salmonella</i> Typhimurium ArtAB Toxin and Its Receptor-Binding Subunit |
title_fullStr | Cellular Activity of <i>Salmonella</i> Typhimurium ArtAB Toxin and Its Receptor-Binding Subunit |
title_full_unstemmed | Cellular Activity of <i>Salmonella</i> Typhimurium ArtAB Toxin and Its Receptor-Binding Subunit |
title_short | Cellular Activity of <i>Salmonella</i> Typhimurium ArtAB Toxin and Its Receptor-Binding Subunit |
title_sort | cellular activity of i salmonella i typhimurium artab toxin and its receptor binding subunit |
topic | <i>Salmonella</i> Typhimurium AB<sub>5</sub> toxin ArtAB |
url | https://www.mdpi.com/2072-6651/13/9/599 |
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