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. 2012 Sep 25;10(1):57.
doi: 10.1186/1477-5956-10-57.

Proteomic identification of MYC2-dependent jasmonate-regulated proteins in Arabidopsis thaliana

Affiliations

Proteomic identification of MYC2-dependent jasmonate-regulated proteins in Arabidopsis thaliana

Jing Guo et al. Proteome Sci. .

Abstract

Background: MYC2, a basic helix-loop-helix (bHLH) domain-containing transcription factor, participates in the jasmonate (JA) signaling pathway and is involved in the modulation of diverse JA functions. However, a comprehensive list of MYC2-dependent JA-responsive proteins has yet to be defined.

Results: In this paper, we report the comparative proteomics of wild-type (WT) plants and jin1-9, a MYC2 mutant plant, in response to methyl jasmonate (MeJA) treatment. Proteins from mock/MeJA-treated jin1-9 and WT samples were extracted and separated by two-dimensional gel electrophoresis. Twenty-seven JA-mediated proteins demonstrated differential expression modulated by MYC2. We observed that MYC2 negatively regulates the accumulation of JA-dependent indolic glucosinolate-related proteins and exhibits opposite effects on the biosynthetic enzymes involved aliphatic glucosinolate pathways. In addition, proteins involved in the tricarboxylic acid cycle and a majority of the MeJA-inducible proteins that are involved in multiple protective systems against oxidative stress were reduced in jin1-9/myc2 sample compared to the WT sample. These results support a positive role for MYC2 in regulating JA-mediated carbohydrate metabolism and oxidative stress tolerance.

Conclusions: We have identified MYC2-dependent jasmonate-regulated proteins in Arabidopsis thaliana by performing two-dimensional gel electrophoresis and MALDI-TOF/TOF MS analysis. The observed pattern of protein expression suggests that MYC2 has opposite effects on the biosynthetic enzymes of indolic and aliphatic glucosinolate pathways and positively regulates JA-mediated carbohydrate metabolism and oxidative stress tolerance-related proteins. Furthermore, it is very interesting to note that MYC2 plays opposite roles in the modulation of a subset of JA-regulated photosynthetic proteins during short-term and long-term JA signaling. This study will enhance our understanding of the function of MYC2 in JA signaling in Arabidopsis thaliana.

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Figures

Figure 1
Figure 1
2-DE gel analysis of proteins extracted from the jin1-9/myc2 mutant. Separation on the first dimension was performed using 1300 μg of total soluble proteins using the linear gradient IPG strips with pH 4–7. In the second dimension, 12.5% SDS-PAGE gels were used, and proteins were visualized using coomassie brilliant blue. Arrows indicate 27 protein spots that changed reproducibly and significantly in MeJA-treated jin1-9/myc2 mutant plants compared to MeJA-treated wild-type plants. A and B represent 2-DE gels from jin1-9/myc2 samples treated with 200 mM MeJA for 6 h and 48 h, respectively. 2-DE experiments were repeated 3 times with independent biological replicates.
Figure 2
Figure 2
Classification of the identified proteins based on their molecular functions. The pie chart indicates the percent distribution of the MYC2-dependent JA-regulated proteins into their functional classes.
Figure 3
Figure 3
Quantitative real-time PCR analysis of the transcription of genes encoding proteins identified in 2-DE. Data are expressed as relative RNA levels ([mRNA]gene/[mRNA]actin) and are the mean of three biological replicates (> 6 pooled plants each); error bars denote SE. Double stars indicate p < 0.01 between MeJA-treated jin1-9/myc2 and wild-type samples.
Figure 4
Figure 4
MYC2 differentially regulates different JA-dependent biological process. The role of MYC2 is indicated in blue and red to specify 6 h and 48 h MeJA treatments, respectively. Arrows and blunt arrows indicate positive and negative regulation, respectively.

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