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Quantitative Proteomic Analysis of Brucella abortus under Multiple Environmental Stresses

ABSTRACT: We have used a multiple-environmental-stress strategy to reveal global metabolic adaptations of B. abortus to intravacuolar environmental conditions. These conditions included: (i) a Control condition (growth on TSB, condition #1); (ii) seven single-stress conditions: (condition #2-8) and (iii) a multi-stress condition (#9). The multi-stress was a combination of each single-stress that may better simulated the environments that Brucella may occur during the infection of a host. The proteomic analysis identified and quantified in total 2,272 proteins and 74% of the theoretical proteome, which is providing a wide coverage of B. abortus genome. The identification of proteins necessary for stress resistance is crucial to elucidate the infectious process in order to control brucellosis and may provide valuable clues towards discovery of novel therapeutic targets and effective vaccines.

ORGANISM(S): Brucella Abortus

SUBMITTER: Xiaodong Zai

PROVIDER: PXD007548 | JPOST Repository | Thu Aug 24 00:00:00 BST 2017

REPOSITORIES: jPOST

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Project description:Organisms frequently experience environmental stresses that occur in predictable patterns and combinations. For wild Saccharomyces cerevisiae yeast growing in natural environments, cells may experience high osmotic stress when they first enter broken fruit, followed by high ethanol levels during fermentation, and then finally high levels of oxidative stress resulting from respiration of ethanol. Yeast have adapted to these patterns by evolving sophisticated “cross protection” mechanisms, where mild ‘primary’ doses of one stress can enhance tolerance to severe doses of a different ‘secondary’ stress. For example, in many yeast strains, mild osmotic or mild ethanol stresses cross protect against severe oxidative stress, which likely reflects an anticipatory response important for high fitness in nature. During the course of genetic mapping studies to understand the mechanisms underlying natural variation in ethanol-induced cross protection against H2O2, we found that a key H2O2 scavenging enzyme, cytosolic catalase T (Ctt1p), was absolutely essential for cross protection in a wild oak strain, suggesting the absence of compensatory mechanisms for acquired H2O2 resistance under that condition. In this study, we found surprising heterogeneity across diverse yeast strains in whether CTT1 function was fully necessary for acquired H2O2 resistance. Some strains exhibited partial dispensability of CTT1 when ethanol and/or salt were used as mild stressors, suggesting that compensatory peroxidases may play a role in acquired stress resistance in certain genetic backgrounds. We leveraged global transcriptional responses to ethanol and salt stresses in strains with different levels of CTT1 dispensability, allowing us to identify possible regulators of these alternative peroxidases and acquired stress resistance in general. Ultimately, this study highlights how superficially similar traits can have different underlying molecular foundations and provides a framework for understanding the diversity and regulation of stress defense mechanisms.

Project description:Considering global climate changes, incidences of combined drought and heat stress are likely to increase in the future and will considerably influence plant-pathogen interactions. Until now, little is known about plants exposed to simultaneously occurring abiotic and biotic stresses. To shed some light on molecular plant responses to multiple stress factors, a versatile multi-factorial test system, allowing simultaneous application of heat, drought and virus stress, was developed. Comparative analysis of single, double and triple stress responses by transcriptome and metabolome analysis revealed that gene expression under multi-factorial stress is not predictable from single stress treatments. Hierarchical cluster and principal component analysis identified heat as the major stress factor clearly separating heat-stressed from non-heat stressed plants. We identified 11 genes differentially regulated in all stress combinations as well as 23 genes specifically-regulated under triple stress. Furthermore, we showed that virus treated plants displayed enhanced expression of defense genes, which was abolished in plants additionally subjected to heat and drought stress. Triple stress also reduced expression of genes involved in the R-mediated disease response and increased the cytoplasmic protein response which was not seen under single stress conditions. These observations suggested that abiotic stress factors significantly altered TuMV-specific signaling networks which lead to a deactivation of defense responses and a higher susceptibility of plants. Collectively, our transcriptome and metabolome data provide a powerful resource to study plant responses during multi-factorial stress and allows identifying metabolic processes and functional networks involved in tripartite interactions of plants with their environment. Stress induced gene expression in Arabidopsis leaves was measured after exposure to single and combined abiotic and biotic stress. Plants were grown on soil for 21 days till virus infection. Eight days later controlled drought stress was applied. At the end of the treatments heat was applied for three days. In parallel, a homozougus T-DNA insertion line SALK_021115C (N672283), located in the Arabidopsis gene At5g45000 has been exposed to the same stress conditions. Four biological replicates have been hybridized for each treatment.

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Quantitative Proteomic Analysis of Brucella abortus under Multiple Environmental Stresses

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