Quantitative proteomic analysis of G-protein signalling in Stagonospora nodorum using isobaric tags for relative and absolute quantification

The G protein α-subunit (Gna1) in the wheat pathogen Stagonospora nodorum has previously been shown to be a critical controlling element in disease ontogeny. In this study, iTRAQ and 2-D LC MALDI-MS/MS have been used to characterise protein expression changes in the S. nodorum gna1 strain versus the...

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Main Authors: Casey, T., Solomon, P., Bringans, S., Tan, Kar-Chun, Oliver, Richard, Lipscombe, R.
Format: Journal Article
Published: 2010
Online Access:http://hdl.handle.net/20.500.11937/11305
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author Casey, T.
Solomon, P.
Bringans, S.
Tan, Kar-Chun
Oliver, Richard
Lipscombe, R.
author_facet Casey, T.
Solomon, P.
Bringans, S.
Tan, Kar-Chun
Oliver, Richard
Lipscombe, R.
author_sort Casey, T.
building Curtin Institutional Repository
collection Online Access
description The G protein α-subunit (Gna1) in the wheat pathogen Stagonospora nodorum has previously been shown to be a critical controlling element in disease ontogeny. In this study, iTRAQ and 2-D LC MALDI-MS/MS have been used to characterise protein expression changes in the S. nodorum gna1 strain versus the SN15 wild-type. A total of 1336 proteins were identified. The abundance of 49 proteins was significantly altered in the gna1 strain compared with the wild-type. Gna1 was identified as having a significant regulatory role on primary metabolic pathways, particularly those concerned with NADPH synthesis or consumption. Mannitol dehydrogenase was up-regulated in the gna1 strain while mannitol 1-phosphate dehydrogenase was down-regulated providing direct evidence of Gna1 regulation over this enigmatic pathway. Enzymatic analysis and growth assays confirmed this regulatory role. Several novel hypothetical proteins previously associated with stress and pathogen responses were identified as positively regulated by Gna1. A short-chain dehydrogenase (Sch3) was also significantly less abundant in the gna1 strains. Sch3 was further characterised by gene disruption in S. nodorum by homologous recombination. Functional characterisation of the sch3 strains revealed their inability to sporulate in planta providing a further link to Gna1 signalling and asexual reproduction. These data add significantly to the identification of the regulatory targets of Gna1 signalling in S. nodorum and have demonstrated the utility of iTRAQ in dissecting signal transduction pathways.
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spelling curtin-20.500.11937-113052017-09-13T16:07:07Z Quantitative proteomic analysis of G-protein signalling in Stagonospora nodorum using isobaric tags for relative and absolute quantification Casey, T. Solomon, P. Bringans, S. Tan, Kar-Chun Oliver, Richard Lipscombe, R. The G protein α-subunit (Gna1) in the wheat pathogen Stagonospora nodorum has previously been shown to be a critical controlling element in disease ontogeny. In this study, iTRAQ and 2-D LC MALDI-MS/MS have been used to characterise protein expression changes in the S. nodorum gna1 strain versus the SN15 wild-type. A total of 1336 proteins were identified. The abundance of 49 proteins was significantly altered in the gna1 strain compared with the wild-type. Gna1 was identified as having a significant regulatory role on primary metabolic pathways, particularly those concerned with NADPH synthesis or consumption. Mannitol dehydrogenase was up-regulated in the gna1 strain while mannitol 1-phosphate dehydrogenase was down-regulated providing direct evidence of Gna1 regulation over this enigmatic pathway. Enzymatic analysis and growth assays confirmed this regulatory role. Several novel hypothetical proteins previously associated with stress and pathogen responses were identified as positively regulated by Gna1. A short-chain dehydrogenase (Sch3) was also significantly less abundant in the gna1 strains. Sch3 was further characterised by gene disruption in S. nodorum by homologous recombination. Functional characterisation of the sch3 strains revealed their inability to sporulate in planta providing a further link to Gna1 signalling and asexual reproduction. These data add significantly to the identification of the regulatory targets of Gna1 signalling in S. nodorum and have demonstrated the utility of iTRAQ in dissecting signal transduction pathways. 2010 Journal Article http://hdl.handle.net/20.500.11937/11305 10.1002/pmic.200900474 restricted
spellingShingle Casey, T.
Solomon, P.
Bringans, S.
Tan, Kar-Chun
Oliver, Richard
Lipscombe, R.
Quantitative proteomic analysis of G-protein signalling in Stagonospora nodorum using isobaric tags for relative and absolute quantification
title Quantitative proteomic analysis of G-protein signalling in Stagonospora nodorum using isobaric tags for relative and absolute quantification
title_full Quantitative proteomic analysis of G-protein signalling in Stagonospora nodorum using isobaric tags for relative and absolute quantification
title_fullStr Quantitative proteomic analysis of G-protein signalling in Stagonospora nodorum using isobaric tags for relative and absolute quantification
title_full_unstemmed Quantitative proteomic analysis of G-protein signalling in Stagonospora nodorum using isobaric tags for relative and absolute quantification
title_short Quantitative proteomic analysis of G-protein signalling in Stagonospora nodorum using isobaric tags for relative and absolute quantification
title_sort quantitative proteomic analysis of g-protein signalling in stagonospora nodorum using isobaric tags for relative and absolute quantification
url http://hdl.handle.net/20.500.11937/11305