A proteomic approach to investigating gene cluster expression and secondary metabolite functionality in Aspergillus fumigatus.

A combined proteomics and metabolomics approach was utilised to advance the identification and characterisation of secondary metabolites in Aspergillus fumigatus. Here, implementation of a shotgun proteomic strategy led to the identification of non-redundant mycelial proteins (n = 414) from A. fumig...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Rebecca A Owens, Stephen Hammel, Kevin J Sheridan, Gary W Jones, Sean Doyle
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/279820b2d9214da3b2aa7e54b4763c22
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:279820b2d9214da3b2aa7e54b4763c22
record_format	dspace
spelling	oai:doaj.org-article:279820b2d9214da3b2aa7e54b4763c222021-11-25T06:01:31ZA proteomic approach to investigating gene cluster expression and secondary metabolite functionality in Aspergillus fumigatus.1932-620310.1371/journal.pone.0106942https://doaj.org/article/279820b2d9214da3b2aa7e54b4763c222014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/25198175/?tool=EBIhttps://doaj.org/toc/1932-6203A combined proteomics and metabolomics approach was utilised to advance the identification and characterisation of secondary metabolites in Aspergillus fumigatus. Here, implementation of a shotgun proteomic strategy led to the identification of non-redundant mycelial proteins (n = 414) from A. fumigatus including proteins typically under-represented in 2-D proteome maps: proteins with multiple transmembrane regions, hydrophobic proteins and proteins with extremes of molecular mass and pI. Indirect identification of secondary metabolite cluster expression was also achieved, with proteins (n = 18) from LaeA-regulated clusters detected, including GliT encoded within the gliotoxin biosynthetic cluster. Biochemical analysis then revealed that gliotoxin significantly attenuates H2O2-induced oxidative stress in A. fumigatus (p>0.0001), confirming observations from proteomics data. A complementary 2-D/LC-MS/MS approach further elucidated significantly increased abundance (p<0.05) of proliferating cell nuclear antigen (PCNA), NADH-quinone oxidoreductase and the gliotoxin oxidoreductase GliT, along with significantly attenuated abundance (p<0.05) of a heat shock protein, an oxidative stress protein and an autolysis-associated chitinase, when gliotoxin and H2O2 were present, compared to H2O2 alone. Moreover, gliotoxin exposure significantly reduced the abundance of selected proteins (p<0.05) involved in de novo purine biosynthesis. Significantly elevated abundance (p<0.05) of a key enzyme, xanthine-guanine phosphoribosyl transferase Xpt1, utilised in purine salvage, was observed in the presence of H2O2 and gliotoxin. This work provides new insights into the A. fumigatus proteome and experimental strategies, plus mechanistic data pertaining to gliotoxin functionality in the organism.Rebecca A OwensStephen HammelKevin J SheridanGary W JonesSean DoylePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 9, p e106942 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rebecca A Owens Stephen Hammel Kevin J Sheridan Gary W Jones Sean Doyle A proteomic approach to investigating gene cluster expression and secondary metabolite functionality in Aspergillus fumigatus.
description	A combined proteomics and metabolomics approach was utilised to advance the identification and characterisation of secondary metabolites in Aspergillus fumigatus. Here, implementation of a shotgun proteomic strategy led to the identification of non-redundant mycelial proteins (n = 414) from A. fumigatus including proteins typically under-represented in 2-D proteome maps: proteins with multiple transmembrane regions, hydrophobic proteins and proteins with extremes of molecular mass and pI. Indirect identification of secondary metabolite cluster expression was also achieved, with proteins (n = 18) from LaeA-regulated clusters detected, including GliT encoded within the gliotoxin biosynthetic cluster. Biochemical analysis then revealed that gliotoxin significantly attenuates H2O2-induced oxidative stress in A. fumigatus (p>0.0001), confirming observations from proteomics data. A complementary 2-D/LC-MS/MS approach further elucidated significantly increased abundance (p<0.05) of proliferating cell nuclear antigen (PCNA), NADH-quinone oxidoreductase and the gliotoxin oxidoreductase GliT, along with significantly attenuated abundance (p<0.05) of a heat shock protein, an oxidative stress protein and an autolysis-associated chitinase, when gliotoxin and H2O2 were present, compared to H2O2 alone. Moreover, gliotoxin exposure significantly reduced the abundance of selected proteins (p<0.05) involved in de novo purine biosynthesis. Significantly elevated abundance (p<0.05) of a key enzyme, xanthine-guanine phosphoribosyl transferase Xpt1, utilised in purine salvage, was observed in the presence of H2O2 and gliotoxin. This work provides new insights into the A. fumigatus proteome and experimental strategies, plus mechanistic data pertaining to gliotoxin functionality in the organism.
format	article
author	Rebecca A Owens Stephen Hammel Kevin J Sheridan Gary W Jones Sean Doyle
author_facet	Rebecca A Owens Stephen Hammel Kevin J Sheridan Gary W Jones Sean Doyle
author_sort	Rebecca A Owens
title	A proteomic approach to investigating gene cluster expression and secondary metabolite functionality in Aspergillus fumigatus.
title_short	A proteomic approach to investigating gene cluster expression and secondary metabolite functionality in Aspergillus fumigatus.
title_full	A proteomic approach to investigating gene cluster expression and secondary metabolite functionality in Aspergillus fumigatus.
title_fullStr	A proteomic approach to investigating gene cluster expression and secondary metabolite functionality in Aspergillus fumigatus.
title_full_unstemmed	A proteomic approach to investigating gene cluster expression and secondary metabolite functionality in Aspergillus fumigatus.
title_sort	proteomic approach to investigating gene cluster expression and secondary metabolite functionality in aspergillus fumigatus.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/279820b2d9214da3b2aa7e54b4763c22
work_keys_str_mv	AT rebeccaaowens aproteomicapproachtoinvestigatinggeneclusterexpressionandsecondarymetabolitefunctionalityinaspergillusfumigatus AT stephenhammel aproteomicapproachtoinvestigatinggeneclusterexpressionandsecondarymetabolitefunctionalityinaspergillusfumigatus AT kevinjsheridan aproteomicapproachtoinvestigatinggeneclusterexpressionandsecondarymetabolitefunctionalityinaspergillusfumigatus AT garywjones aproteomicapproachtoinvestigatinggeneclusterexpressionandsecondarymetabolitefunctionalityinaspergillusfumigatus AT seandoyle aproteomicapproachtoinvestigatinggeneclusterexpressionandsecondarymetabolitefunctionalityinaspergillusfumigatus AT rebeccaaowens proteomicapproachtoinvestigatinggeneclusterexpressionandsecondarymetabolitefunctionalityinaspergillusfumigatus AT stephenhammel proteomicapproachtoinvestigatinggeneclusterexpressionandsecondarymetabolitefunctionalityinaspergillusfumigatus AT kevinjsheridan proteomicapproachtoinvestigatinggeneclusterexpressionandsecondarymetabolitefunctionalityinaspergillusfumigatus AT garywjones proteomicapproachtoinvestigatinggeneclusterexpressionandsecondarymetabolitefunctionalityinaspergillusfumigatus AT seandoyle proteomicapproachtoinvestigatinggeneclusterexpressionandsecondarymetabolitefunctionalityinaspergillusfumigatus
_version_	1718414270601887744

A proteomic approach to investigating gene cluster expression and secondary metabolite functionality in Aspergillus fumigatus.

Ejemplares similares