Proteomic and phosphoproteomic approaches to understand plant–pathogen interactions

Thurston, Graham S.; Regan, Sharon; Rampitsch, Chris; Xing, Tim

doi:10.1016/j.pmpp.2005.03.004

Cited by 41 publications

(32 citation statements)

References 49 publications

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“…It appears to be the most predominant PTM in plants in response to pathogens (5). Many signaling components, such as protein kinases, phosphatases, and transcription factors, have been implied in relation to changes of phosphorylation status during plant-pathogen interactions (5).…”

mentioning

confidence: 99%

Battle through Signaling between Wheat and the Fungal Pathogen Septoria tritici Revealed by Proteomics and Phosphoproteomics

Yang

Melo‐Braga

Larsen

et al. 2013

Molecular & Cellular Proteomics

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The fungus Septoria tritici causes the disease septoria tritici blotch in wheat, one of the most economically devastating foliar diseases in this crop. To investigate signaling events and defense responses in the wheat-S. tritici interaction, we performed a time-course study of S. tritici infection in resistant and susceptible wheat using quantitative proteomics and phosphoproteomics, with special emphasis on the initial biotrophic phase of interactions. Our study revealed an accumulation of defense and stress-related proteins, suppression of photosynthesis, and changes in sugar metabolism during compatible and incompatible interactions. However, differential regulation of the phosphorylation status of signaling proteins, transcription and translation regulators, and membraneassociated proteins was observed between two interactions. The proteomic data were correlated with a more rapid or stronger accumulation of signal molecules, including calcium, H 2 O 2 , NO, and sugars, in the resistant than in the susceptible cultivar in response to the infection. Additionally, 31 proteins and 5 phosphoproteins from the pathogen were identified, including metabolic proteins and signaling proteins such as GTP-binding proteins, 14 -3-3 proteins, and calcium-binding proteins. Quantitative PCR analysis showed the expression of fungal signaling genes and genes encoding a superoxide dismutase and cell-wall degrading enzymes. These results indicate roles of signaling, antioxidative stress mechanisms, and nutrient acquisition in facilitating the initial symptomless growth. Taken in its entirety, our dataset suggests interplay between the plant and S. tritici through complex signaling networks and downstream molecular events. Resistance is likely related to several rapidly and intensively triggered signal transduction cascades resulting in a multiple-level activation of transcription and translation processes of defense responses. Our sensitive approaches and model provide a comprehensive (phospho)proteomics resource for studying signaling from the point of view of both host and pathogen during a plant-pathogen interaction. Molecular & Cellular Proteomics 12:

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mentioning

confidence: 99%

Battle through Signaling between Wheat and the Fungal Pathogen Septoria tritici Revealed by Proteomics and Phosphoproteomics

Yang

Melo‐Braga

Larsen

et al. 2013

Molecular & Cellular Proteomics

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“…3 As indicated by Benschop et al among the early responding proteins following an elicitor stimulation in Arabidopsis, a substantial number (12 of 76) of the differentially phosphorylated proteins were enzymes involved in protein phosphorylation and dephosphorylation, including different families of kinases (3 calciumdependent protein kinases, CDPKs), receptor-like kinase (RLKs) and protein phosphatases. Using IMAC and MS, a large scale analysis of phosphorylated membrane proteins was performed, phosphopeptides from several isoforms of PM H + -ATPase were identified, and more importantly, novel phosphorylation sites in these isoforms were found.…”

Section: Plasma Membrane H + -Atpasementioning

confidence: 99%

“…It is also true that some early responses may involve only post-translational regulation. 3 Proteomics has mostly been used to seek out over or underexpression of proteins separated by two-dimensional PAGE (2DE) by isoelectric point and molecular mass. The 2DE can be used to directly detect phosphorylated proteins using specific stain (e.g., Pro-Q Diamond staining) or by a more sensitive methods based on immunodetection of phosphorylated proteins after protein gel blotting to analyze their post-translational modifications.…”

Section: 3mentioning

confidence: 99%

“…In an activated tomato tMEK2 pathway, a phosphoproteomics screening failed to find some known downstream kinases 3,26 such as LeMPK3, which was later identified in an in vitro protein interaction analysis. 26 While the abundance of the kinases or their substrates is one of the key limiting elements in detection, a transient nature of kinase function should be considered carefully.…”

Section: Prospects In Determining the Precise Mechanisms Of Phosphorymentioning

confidence: 99%

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Revealing plant defense signaling

Xing¹,

Laroche²

2011

Plant Signaling & Behavior

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“…A growing body of evidence highlights the importance of PTMs in different aspects of plant immunity 1,5,6 . Protein phosphorylation, the reversible attachment of a phosphate group to a serine, threonine or tyrosine residue is a regulator of many cellular functions and not surprisingly the most highly studied PTM in plant defense signaling cascades [5][6][7][8][9][10][11] . Phosphorylationdependent signaling is an integral part of plant defense activation initiated after extracellular perception of microbes by transmembrane receptors, or intracellular recognition by multidomain resistance proteins 5,8 .…”

Section: Introductionmentioning

confidence: 99%

Identification of Post-translational Modifications of Plant Protein Complexes

Piquerez

Balmuth

Sklenář

et al. 2014

JoVE

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Plants adapt quickly to changing environments due to elaborate perception and signaling systems. During pathogen attack, plants rapidly respond to infection via the recruitment and activation of immune complexes. Activation of immune complexes is associated with posttranslational modifications (PTMs) of proteins, such as phosphorylation, glycosylation, or ubiquitination. Understanding how these PTMs are choreographed will lead to a better understanding of how resistance is achieved.Here we describe a protein purification method for nucleotide-binding leucine-rich repeat (NB-LRR)-interacting proteins and the subsequent identification of their post-translational modifications (PTMs). With small modifications, the protocol can be applied for the purification of other plant protein complexes. The method is based on the expression of an epitope-tagged version of the protein of interest, which is subsequently partially purified by immunoprecipitation and subjected to mass spectrometry for identification of interacting proteins and PTMs. This protocol demonstrates that: i). Dynamic changes in PTMs such as phosphorylation can be detected by mass spectrometry; ii). It is important to have sufficient quantities of the protein of interest, and this can compensate for the lack of purity of the immunoprecipitate; iii). In order to detect PTMs of a protein of interest, this protein has to be immunoprecipitated to get a sufficient quantity of protein.

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Proteomic and phosphoproteomic approaches to understand plant–pathogen interactions

Cited by 41 publications

References 49 publications

Battle through Signaling between Wheat and the Fungal Pathogen Septoria tritici Revealed by Proteomics and Phosphoproteomics

Battle through Signaling between Wheat and the Fungal Pathogen Septoria tritici Revealed by Proteomics and Phosphoproteomics

Revealing plant defense signaling

Identification of Post-translational Modifications of Plant Protein Complexes

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