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Proteome dynamics and early salt stress response of the photosynthetic organism Chlamydomonas reinhardtii

BACKGROUND: The cellular proteome and metabolome are underlying dynamic regulation allowing rapid adaptation to changes in the environment. System-wide analysis of these dynamics will provide novel insights into mechanisms of stress adaptation for higher photosynthetic organisms. We applied pulsed-S...

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Autores principales: Mastrobuoni, Guido, Irgang, Susann, Pietzke, Matthias, Aßmus, Heike E, Wenzel, Markus, Schulze, Waltraud X, Kempa, Stefan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3444938/
https://www.ncbi.nlm.nih.gov/pubmed/22651860
http://dx.doi.org/10.1186/1471-2164-13-215
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author Mastrobuoni, Guido
Irgang, Susann
Pietzke, Matthias
Aßmus, Heike E
Wenzel, Markus
Schulze, Waltraud X
Kempa, Stefan
author_facet Mastrobuoni, Guido
Irgang, Susann
Pietzke, Matthias
Aßmus, Heike E
Wenzel, Markus
Schulze, Waltraud X
Kempa, Stefan
author_sort Mastrobuoni, Guido
collection PubMed
description BACKGROUND: The cellular proteome and metabolome are underlying dynamic regulation allowing rapid adaptation to changes in the environment. System-wide analysis of these dynamics will provide novel insights into mechanisms of stress adaptation for higher photosynthetic organisms. We applied pulsed-SILAC labeling to a photosynthetic organism for the first time and we established a method to study proteome dynamics in the green alga Chlamydomonas reinhardtii, an emerging model system for plant biology. In addition, we combined the analysis of protein synthesis with metabolic profiling to study the dynamic changes of metabolism and proteome turnover under salt stress conditions. RESULTS: To study de novo protein synthesis an arginine auxotroph Chlamydomonas strain was cultivated in presence of stable isotope-labeled arginine for 24 hours. From the time course experiment in 3 salt concentrations we could identify more than 2500 proteins and their H/L ratio in at least one experimental condition; for 998 protiens at least 3 ratio counts were detected in the 24 h time point (0 mM NaCl). After fractionation we could identify 3115 proteins and for 1765 of them we determined their de novo synthesis rate. Consistently with previous findings we showed that RuBisCO is among the most prominent proteins in the cell; and similar abundance and turnover for the small and large RuBisCO subunit could be calculated. The D1 protein was identified among proteins with a high synthesis rates. A global median half-life of 45 h was calculated for Chlamydomonas proteins under the chosen conditions. CONCLUSION: To investigate the temporal co-regulation of the proteome and metabolome, we applied salt stress to Chlamydomonas and studied the time dependent regulation of protein expression and changes in the metabolome. The main metabolic response to salt stress was observed within the amino acid metabolism. In particular, proline was up-regulated manifold and according to that an increased carbon flow within the proline biosynthetic pathway could be measured. In parallel the analysis of abundance and de novo synthesis of the corresponding enzymes revealed that metabolic rearrangements precede adjustments of protein abundance.
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spelling pubmed-34449382012-09-19 Proteome dynamics and early salt stress response of the photosynthetic organism Chlamydomonas reinhardtii Mastrobuoni, Guido Irgang, Susann Pietzke, Matthias Aßmus, Heike E Wenzel, Markus Schulze, Waltraud X Kempa, Stefan BMC Genomics Research Article BACKGROUND: The cellular proteome and metabolome are underlying dynamic regulation allowing rapid adaptation to changes in the environment. System-wide analysis of these dynamics will provide novel insights into mechanisms of stress adaptation for higher photosynthetic organisms. We applied pulsed-SILAC labeling to a photosynthetic organism for the first time and we established a method to study proteome dynamics in the green alga Chlamydomonas reinhardtii, an emerging model system for plant biology. In addition, we combined the analysis of protein synthesis with metabolic profiling to study the dynamic changes of metabolism and proteome turnover under salt stress conditions. RESULTS: To study de novo protein synthesis an arginine auxotroph Chlamydomonas strain was cultivated in presence of stable isotope-labeled arginine for 24 hours. From the time course experiment in 3 salt concentrations we could identify more than 2500 proteins and their H/L ratio in at least one experimental condition; for 998 protiens at least 3 ratio counts were detected in the 24 h time point (0 mM NaCl). After fractionation we could identify 3115 proteins and for 1765 of them we determined their de novo synthesis rate. Consistently with previous findings we showed that RuBisCO is among the most prominent proteins in the cell; and similar abundance and turnover for the small and large RuBisCO subunit could be calculated. The D1 protein was identified among proteins with a high synthesis rates. A global median half-life of 45 h was calculated for Chlamydomonas proteins under the chosen conditions. CONCLUSION: To investigate the temporal co-regulation of the proteome and metabolome, we applied salt stress to Chlamydomonas and studied the time dependent regulation of protein expression and changes in the metabolome. The main metabolic response to salt stress was observed within the amino acid metabolism. In particular, proline was up-regulated manifold and according to that an increased carbon flow within the proline biosynthetic pathway could be measured. In parallel the analysis of abundance and de novo synthesis of the corresponding enzymes revealed that metabolic rearrangements precede adjustments of protein abundance. BioMed Central 2012-05-31 /pmc/articles/PMC3444938/ /pubmed/22651860 http://dx.doi.org/10.1186/1471-2164-13-215 Text en Copyright ©2012 Mastrobuoni et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Mastrobuoni, Guido
Irgang, Susann
Pietzke, Matthias
Aßmus, Heike E
Wenzel, Markus
Schulze, Waltraud X
Kempa, Stefan
Proteome dynamics and early salt stress response of the photosynthetic organism Chlamydomonas reinhardtii
title Proteome dynamics and early salt stress response of the photosynthetic organism Chlamydomonas reinhardtii
title_full Proteome dynamics and early salt stress response of the photosynthetic organism Chlamydomonas reinhardtii
title_fullStr Proteome dynamics and early salt stress response of the photosynthetic organism Chlamydomonas reinhardtii
title_full_unstemmed Proteome dynamics and early salt stress response of the photosynthetic organism Chlamydomonas reinhardtii
title_short Proteome dynamics and early salt stress response of the photosynthetic organism Chlamydomonas reinhardtii
title_sort proteome dynamics and early salt stress response of the photosynthetic organism chlamydomonas reinhardtii
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3444938/
https://www.ncbi.nlm.nih.gov/pubmed/22651860
http://dx.doi.org/10.1186/1471-2164-13-215
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