Staphylococcus aureus interaction with phospholipid vesicles--a new method to accurately determine accessory gene regulator (agr) activity.

The staphylococcal accessory gene regulatory (agr) operon is a well-characterised global regulatory element that is important in the control of virulence gene expression for Staphylococcus aureus, a major human pathogen. Hence, accurate and sensitive measurement of Agr activity is central in underst...

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Autores principales:	Maisem Laabei, W David Jamieson, Ruth C Massey, A Tobias A Jenkins
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/584d690e4b0144f8aa5b5b8cd51eb855
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spelling	oai:doaj.org-article:584d690e4b0144f8aa5b5b8cd51eb8552021-11-18T08:34:48ZStaphylococcus aureus interaction with phospholipid vesicles--a new method to accurately determine accessory gene regulator (agr) activity.1932-620310.1371/journal.pone.0087270https://doaj.org/article/584d690e4b0144f8aa5b5b8cd51eb8552014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24498061/?tool=EBIhttps://doaj.org/toc/1932-6203The staphylococcal accessory gene regulatory (agr) operon is a well-characterised global regulatory element that is important in the control of virulence gene expression for Staphylococcus aureus, a major human pathogen. Hence, accurate and sensitive measurement of Agr activity is central in understanding the virulence potential of Staphylococcus aureus, especially in the context of Agr dysfunction, which has been linked with persistent bacteraemia and reduced susceptibility to glycopeptide antibiotics. Agr function is typically measured using a synergistic haemolysis CAMP assay, which is believe to report on the level of expression of one of the translated products of the agr locus, delta toxin. In this study we develop a vesicle lysis test (VLT) that is specific to small amphipathic peptides, most notably delta and Phenol Soluble Modulin (PSM) toxins. To determine the accuracy of this VLT method in assaying Agr activity, we compared it to the CAMP assay using 89 clinical Staphylococcus aureus isolates. Of the 89 isolates, 16 were designated as having dysfunctional Agr systems by the CAMP assay, whereas only three were designated as such by VLT. Molecular analysis demonstrated that of these 16 isolates, the 13 designated as having a functional Agr system by VLT transcribed rnaIII and secreted delta toxin, demonstrating they have a functional Agr system despite the results of the CAMP assay. The agr locus of all 16 isolates was sequenced, and only the 3 designated as having a dysfunctional Agr system contained mutations, explaining their Agr dysfunction. Given the potentially important link between Agr dysfunction and clinical outcome, we have developed an assay that determines this more accurately than the conventional CAMP assay.Maisem LaabeiW David JamiesonRuth C MasseyA Tobias A JenkinsPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 1, p e87270 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Maisem Laabei W David Jamieson Ruth C Massey A Tobias A Jenkins Staphylococcus aureus interaction with phospholipid vesicles--a new method to accurately determine accessory gene regulator (agr) activity.
description	The staphylococcal accessory gene regulatory (agr) operon is a well-characterised global regulatory element that is important in the control of virulence gene expression for Staphylococcus aureus, a major human pathogen. Hence, accurate and sensitive measurement of Agr activity is central in understanding the virulence potential of Staphylococcus aureus, especially in the context of Agr dysfunction, which has been linked with persistent bacteraemia and reduced susceptibility to glycopeptide antibiotics. Agr function is typically measured using a synergistic haemolysis CAMP assay, which is believe to report on the level of expression of one of the translated products of the agr locus, delta toxin. In this study we develop a vesicle lysis test (VLT) that is specific to small amphipathic peptides, most notably delta and Phenol Soluble Modulin (PSM) toxins. To determine the accuracy of this VLT method in assaying Agr activity, we compared it to the CAMP assay using 89 clinical Staphylococcus aureus isolates. Of the 89 isolates, 16 were designated as having dysfunctional Agr systems by the CAMP assay, whereas only three were designated as such by VLT. Molecular analysis demonstrated that of these 16 isolates, the 13 designated as having a functional Agr system by VLT transcribed rnaIII and secreted delta toxin, demonstrating they have a functional Agr system despite the results of the CAMP assay. The agr locus of all 16 isolates was sequenced, and only the 3 designated as having a dysfunctional Agr system contained mutations, explaining their Agr dysfunction. Given the potentially important link between Agr dysfunction and clinical outcome, we have developed an assay that determines this more accurately than the conventional CAMP assay.
format	article
author	Maisem Laabei W David Jamieson Ruth C Massey A Tobias A Jenkins
author_facet	Maisem Laabei W David Jamieson Ruth C Massey A Tobias A Jenkins
author_sort	Maisem Laabei
title	Staphylococcus aureus interaction with phospholipid vesicles--a new method to accurately determine accessory gene regulator (agr) activity.
title_short	Staphylococcus aureus interaction with phospholipid vesicles--a new method to accurately determine accessory gene regulator (agr) activity.
title_full	Staphylococcus aureus interaction with phospholipid vesicles--a new method to accurately determine accessory gene regulator (agr) activity.
title_fullStr	Staphylococcus aureus interaction with phospholipid vesicles--a new method to accurately determine accessory gene regulator (agr) activity.
title_full_unstemmed	Staphylococcus aureus interaction with phospholipid vesicles--a new method to accurately determine accessory gene regulator (agr) activity.
title_sort	staphylococcus aureus interaction with phospholipid vesicles--a new method to accurately determine accessory gene regulator (agr) activity.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/584d690e4b0144f8aa5b5b8cd51eb855
work_keys_str_mv	AT maisemlaabei staphylococcusaureusinteractionwithphospholipidvesiclesanewmethodtoaccuratelydetermineaccessorygeneregulatoragractivity AT wdavidjamieson staphylococcusaureusinteractionwithphospholipidvesiclesanewmethodtoaccuratelydetermineaccessorygeneregulatoragractivity AT ruthcmassey staphylococcusaureusinteractionwithphospholipidvesiclesanewmethodtoaccuratelydetermineaccessorygeneregulatoragractivity AT atobiasajenkins staphylococcusaureusinteractionwithphospholipidvesiclesanewmethodtoaccuratelydetermineaccessorygeneregulatoragractivity
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Staphylococcus aureus interaction with phospholipid vesicles--a new method to accurately determine accessory gene regulator (agr) activity.

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