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Redox proteomics of tomato in response to Pseudomonas syringae infection.

Balmant KM, Parker J, Yoo MJ, Zhu N, Dufresne C, Chen S - Hortic Res (2015)

Bottom Line: In addition, the results of the redox changes were compared and corrected with the protein level changes.A total of 90 potential redox-regulated proteins were identified with functions in carbohydrate and energy metabolism, biosynthesis of cysteine, sucrose and brassinosteroid, cell wall biogenesis, polysaccharide/starch biosynthesis, cuticle development, lipid metabolism, proteolysis, tricarboxylic acid cycle, protein targeting to vacuole, and oxidation-reduction.This inventory of previously unknown protein redox switches in tomato pathogen defense lays a foundation for future research toward understanding the biological significance of protein redox modifications in plant defense responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Genetics Institute, University of Florida , Gainesville, FL, USA ; Plant Molecular and Cellular Biology Program, University of Florida , Gainesville, FL, USA.

ABSTRACT
Unlike mammals with adaptive immunity, plants rely on their innate immunity based on pattern-triggered immunity (PTI) and effector-triggered immunity (ETI) for pathogen defense. Reactive oxygen species, known to play crucial roles in PTI and ETI, can perturb cellular redox homeostasis and lead to changes of redox-sensitive proteins through modification of cysteine sulfhydryl groups. Although redox regulation of protein functions has emerged as an important mechanism in several biological processes, little is known about redox proteins and how they function in PTI and ETI. In this study, cysTMT proteomics technology was used to identify similarities and differences of protein redox modifications in tomato resistant (PtoR) and susceptible (prf3) genotypes in response to Pseudomonas syringae pv tomato (Pst) infection. In addition, the results of the redox changes were compared and corrected with the protein level changes. A total of 90 potential redox-regulated proteins were identified with functions in carbohydrate and energy metabolism, biosynthesis of cysteine, sucrose and brassinosteroid, cell wall biogenesis, polysaccharide/starch biosynthesis, cuticle development, lipid metabolism, proteolysis, tricarboxylic acid cycle, protein targeting to vacuole, and oxidation-reduction. This inventory of previously unknown protein redox switches in tomato pathogen defense lays a foundation for future research toward understanding the biological significance of protein redox modifications in plant defense responses.

No MeSH data available.


Related in: MedlinePlus

Comparative ontology analysis of the redox proteins after Pst infection between susceptible and resistant genotypes at early and late stages of infection. Relevant biological process GO terms are shown on the y-axis. Numbers of proteins significantly redox-regulated in each genotype and time point after infection are shown on the x-axis.
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fig3: Comparative ontology analysis of the redox proteins after Pst infection between susceptible and resistant genotypes at early and late stages of infection. Relevant biological process GO terms are shown on the y-axis. Numbers of proteins significantly redox-regulated in each genotype and time point after infection are shown on the x-axis.

Mentions: In the susceptible prf3 genotype, 19 cysteine-containing peptides from 19 different proteins were found to be redox-regulated upon pathogen infection (Table 1). Ten of the peptides were found to be in more reduced state at 4 hai with Pst, while the other nine were in more oxidized state compared to the mock control samples (Figure 2). The identification of proteins in a more reduced state under Pst inoculation suggests differential compartmentalization of subcellular redox environment and the dynamic nature of protein redox state. When the total protein-level change and transcriptional change were taken into account, two out of the 19 peptides containing redox-sensitive cysteines also showed significant protein amount changes in the same direction (Table 1). Vacuolar-sorting receptor 1-like and dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex-like were initially assigned as potentially redox-regulated based on the cysTMT data. However, when the protein level correction (based on the iTRAQ data) was applied, these proteins also showed total protein level change. Thus, the observed redox-fold change of these proteins may be due to the protein level change rather than a cysteine redox response. The sequences of the identified redox-regulated proteins were analyzed for intra-molecular disulfide bond formation using DiANNA software (http://clavius.bc.edu/~clotelab/DiANNA/). Five of the 17 redox-sensitive peptides were predicted to form intra-molecular disulfide bonds (Table 1). It should be noted that disulfide bonds represent only one possibility of thiol modifications, and other modifications, such as sulfenic acid, S-nitrosylation and glutathionylation33 are also chemically reversible. The cysTMT labeling method will allow the identification of these cysteine modifications if modification specific reductants are used, e.g., ascorbate and copper for S-nitrosylation.34,35 As shown in Figure 3, analysis of GO terms of relevant biological processes revealed that the potential redox-regulated proteins were involved cellular carbohydrate metabolism, energy processes (plasma membrane adenosine triphosphate (ATPase), ATP synthesis, photosynthesis, and pentose-phosphate shunt), as well as gluconeogenesis and cysteine biosynthetic process. The oxidation–reduction process was represented by five proteins (ferredoxin, peroxidase 12-like, glutamine synthetase (GS), cysteine synthase, and lactoylglutathione lyase-like) in the prf3 genotype inoculated with Pst for 4 h.


Redox proteomics of tomato in response to Pseudomonas syringae infection.

Balmant KM, Parker J, Yoo MJ, Zhu N, Dufresne C, Chen S - Hortic Res (2015)

Comparative ontology analysis of the redox proteins after Pst infection between susceptible and resistant genotypes at early and late stages of infection. Relevant biological process GO terms are shown on the y-axis. Numbers of proteins significantly redox-regulated in each genotype and time point after infection are shown on the x-axis.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591677&req=5

fig3: Comparative ontology analysis of the redox proteins after Pst infection between susceptible and resistant genotypes at early and late stages of infection. Relevant biological process GO terms are shown on the y-axis. Numbers of proteins significantly redox-regulated in each genotype and time point after infection are shown on the x-axis.
Mentions: In the susceptible prf3 genotype, 19 cysteine-containing peptides from 19 different proteins were found to be redox-regulated upon pathogen infection (Table 1). Ten of the peptides were found to be in more reduced state at 4 hai with Pst, while the other nine were in more oxidized state compared to the mock control samples (Figure 2). The identification of proteins in a more reduced state under Pst inoculation suggests differential compartmentalization of subcellular redox environment and the dynamic nature of protein redox state. When the total protein-level change and transcriptional change were taken into account, two out of the 19 peptides containing redox-sensitive cysteines also showed significant protein amount changes in the same direction (Table 1). Vacuolar-sorting receptor 1-like and dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex-like were initially assigned as potentially redox-regulated based on the cysTMT data. However, when the protein level correction (based on the iTRAQ data) was applied, these proteins also showed total protein level change. Thus, the observed redox-fold change of these proteins may be due to the protein level change rather than a cysteine redox response. The sequences of the identified redox-regulated proteins were analyzed for intra-molecular disulfide bond formation using DiANNA software (http://clavius.bc.edu/~clotelab/DiANNA/). Five of the 17 redox-sensitive peptides were predicted to form intra-molecular disulfide bonds (Table 1). It should be noted that disulfide bonds represent only one possibility of thiol modifications, and other modifications, such as sulfenic acid, S-nitrosylation and glutathionylation33 are also chemically reversible. The cysTMT labeling method will allow the identification of these cysteine modifications if modification specific reductants are used, e.g., ascorbate and copper for S-nitrosylation.34,35 As shown in Figure 3, analysis of GO terms of relevant biological processes revealed that the potential redox-regulated proteins were involved cellular carbohydrate metabolism, energy processes (plasma membrane adenosine triphosphate (ATPase), ATP synthesis, photosynthesis, and pentose-phosphate shunt), as well as gluconeogenesis and cysteine biosynthetic process. The oxidation–reduction process was represented by five proteins (ferredoxin, peroxidase 12-like, glutamine synthetase (GS), cysteine synthase, and lactoylglutathione lyase-like) in the prf3 genotype inoculated with Pst for 4 h.

Bottom Line: In addition, the results of the redox changes were compared and corrected with the protein level changes.A total of 90 potential redox-regulated proteins were identified with functions in carbohydrate and energy metabolism, biosynthesis of cysteine, sucrose and brassinosteroid, cell wall biogenesis, polysaccharide/starch biosynthesis, cuticle development, lipid metabolism, proteolysis, tricarboxylic acid cycle, protein targeting to vacuole, and oxidation-reduction.This inventory of previously unknown protein redox switches in tomato pathogen defense lays a foundation for future research toward understanding the biological significance of protein redox modifications in plant defense responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Genetics Institute, University of Florida , Gainesville, FL, USA ; Plant Molecular and Cellular Biology Program, University of Florida , Gainesville, FL, USA.

ABSTRACT
Unlike mammals with adaptive immunity, plants rely on their innate immunity based on pattern-triggered immunity (PTI) and effector-triggered immunity (ETI) for pathogen defense. Reactive oxygen species, known to play crucial roles in PTI and ETI, can perturb cellular redox homeostasis and lead to changes of redox-sensitive proteins through modification of cysteine sulfhydryl groups. Although redox regulation of protein functions has emerged as an important mechanism in several biological processes, little is known about redox proteins and how they function in PTI and ETI. In this study, cysTMT proteomics technology was used to identify similarities and differences of protein redox modifications in tomato resistant (PtoR) and susceptible (prf3) genotypes in response to Pseudomonas syringae pv tomato (Pst) infection. In addition, the results of the redox changes were compared and corrected with the protein level changes. A total of 90 potential redox-regulated proteins were identified with functions in carbohydrate and energy metabolism, biosynthesis of cysteine, sucrose and brassinosteroid, cell wall biogenesis, polysaccharide/starch biosynthesis, cuticle development, lipid metabolism, proteolysis, tricarboxylic acid cycle, protein targeting to vacuole, and oxidation-reduction. This inventory of previously unknown protein redox switches in tomato pathogen defense lays a foundation for future research toward understanding the biological significance of protein redox modifications in plant defense responses.

No MeSH data available.


Related in: MedlinePlus