Higher endogenous methionine in transgenic Arabidopsis seeds affects the composition of storage proteins and lipids

Cohen, Hagai; Pająk, Agnieszka; Pandurangan, Sudhakar; Amir, Rachel; Marsolais, Frédéric

doi:10.1007/s00726-016-2193-4

Cited by 17 publications

(12 citation statements)

References 47 publications

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“…2B), in accordance with previous reports (Matityahu et al, 2013;Cohen et al, 2014Cohen et al, , 2016b. The higher levels of Met and other amino acids in these seeds are apparently due to higher synthesis of several 12S-globulins and 2S-albumin subunits, which are the major seed-storage proteins, as we reported previously for Arabidopsis seeds expressing AtCGS (Cohen et al, 2016a). Despite these lines of evidence, we cannot rule out the possibility that proteins also were degraded, albeit at lower rates than their synthesis, and thus kinetic measurements should be made to examine this notion further.…”

Section: Protein Degradation And/or Decreased Catabolism Of Met Couldsupporting

confidence: 92%

Repression of CYSTATHIONINE γ-SYNTHASE in Seeds Recruits the S-Methylmethionine Cycle

et al. 2017

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-Methylmethionine (SMM) was suggested previously to participate in the metabolism of methionine (Met) in seeds. To further reveal its roles, we had previously produced transgenic Arabidopsis () RNA interference (RNAi) seeds with lower transcript expression of CYSTATHIONINE γ-SYNTHASE (), Met's main regulatory enzyme. Unexpectedly, these seeds accumulated significantly higher levels of Met compared with control seeds through an as yet unknown mechanism. Here, transcript and metabolic analyses coupled with isotope-labeled [C]SMM and [C]Met feeding experiments enabled us to reveal that SMM that was synthesized in rosette leaves of RNAi plants significantly contributed to the accumulation of Met in their seeds at late stages of development. Seed-specific repression of in RNAi seeds triggered the induction of genes operating in the SMM cycle of rosette leaves, leading to elevated transport of SMM toward the seeds, where higher reconversion rates of SMM to Met were detected. The metabolic rearrangements in RNAi seeds resulted in an altered sulfur-associated metabolism, such as lower amounts of Cys and glutathione, as well as a differential composition of glucosinolates. Together, the data propose a novel cross talk existing between seeds and rosette leaves along with mutual effects between the Asp family and SMM pathways operating in these tissues. They also shed light on the effects of higher Met levels on seed physiology and behavior.

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Section: Protein Degradation And/or Decreased Catabolism Of Met Couldsupporting

confidence: 92%

Repression of CYSTATHIONINE γ-SYNTHASE in Seeds Recruits the S-Methylmethionine Cycle

et al. 2017

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“…In addition, the sulfur-containing amino acid, methionine, is known as proteinogenic amino acid and plays a major role in diverse metabolic pathways especially being incorporated into the storage protein synthesis steps [ 58 ]. A higher concentration of methionine led to alteration of the composition of storage proteins, where in particular the 12S-albumins and 2S-globulins presented higher amounts in transgenic Arabidopsis seeds [ 59 ]. In addition, Song et al (2013) demonstrated that transgenic soybean with overexpression of Arabidopsis cystathionine γ-synthase (CGS) showed significant accumulation of methionine than the wild-type seeds and up to a 3.5-fold increase of FAAs in ZD24 cultivar of soybean seeds was seen [ 60 ].…”

Section: Discussionmentioning

confidence: 99%

In-Depth Investigation of Low-Abundance Proteins in Matured and Filling Stages Seeds of Glycine max Employing a Combination of Protamine Sulfate Precipitation and TMT-Based Quantitative Proteomic Analysis

Min

Park

Bae

et al. 2020

Cells

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Despite the significant technical advancements in mass spectrometry-based proteomics and bioinformatics resources, dynamic resolution of soybean seed proteome is still limited because of the high abundance of seed storage proteins (SSPs). These SSPs occupy a large proportion of the total seed protein and hinder the identification of low-abundance proteins. Here, we report a TMT-based quantitative proteome analysis of matured and filling stages seeds of high-protein (Saedanbaek) and low-protein (Daewon) soybean cultivars by application of a two-way pre-fractionation both at the levels of proteins (by PS) and peptides (by basic pH reverse phase chromatography). Interestingly, this approach led to the identification of more than 5900 proteins which is the highest number of proteins reported to date from soybean seeds. Comparative protein profiles of Saedanbaek and Daewon led to the identification of 2200 and 924 differential proteins in mature and filling stages seeds, respectively. Functional annotation of the differential proteins revealed enrichment of proteins related to major metabolism including amino acid, major carbohydrate, and lipid metabolism. In parallel, analysis of free amino acids and fatty acids in the filling stages showed higher contents of all the amino acids in the Saedanbaek while the fatty acids contents were found to be higher in the Daewon. Taken together, these results provide new insights into proteome changes during filling stages in soybean seeds. Moreover, results reported here also provide a framework for systemic and large-scale dissection of seed proteome for the seeds rich in SSPs by two-way pre-fractionation combined with TMT-based quantitative proteome analysis.

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“…Similar observation for that threshold was recently obtained by proteomic profiling of seed-storage proteins in the SSE-AtD-CGS genotype. SSE-AtD-CGS 2 seeds that accumulated sixfold more methionine than the WT exhibited significantly altered composition of its major seed-storage proteins (12S-globulins and 2S-albumins), while no significant changes were detected in the profile of these proteins in SSE-AtD-CGS 1 seeds that accumulated fourfold more methionine than WT seeds (Cohen et al 2016a).…”

Section: Discussionmentioning

confidence: 94%

“…Additionally, Affymetrix microarray analysis employed two selected transgenic Arabidopsis seeds (named SSE-AtD-CGS 1 and 2) that contained four and sixfold more methionine than wild-type (WT) seeds, respectively, further demonstrating that the majority of highly expressed transcripts were related to early desiccation stress responses (Cohen et al 2014). Total protein content was also elevated in these seeds accompanied by altered profiles of seed-storage proteins and fatty acids (Cohen et al 2016a).…”

Section: Electronic Supplementary Materialsmentioning

confidence: 98%

Dose-dependent effects of higher methionine levels on the transcriptome and metabolome of transgenic Arabidopsis seeds

Cohen

Amir

2016

Plant Cell Rep

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Higher methionine levels in transgenic Arabidopsis seeds trigger the accumulation of stress-related transcripts and primary metabolites. These responses depend on the levels of methionine within seeds. Methionine, a sulfur-containing amino acid, is a key metabolite in plant cells. To reveal the regulatory role of the Arabidopsis thaliana CYSTATHIONINE γ-SYNTHASE (AtCGS), methionine main regulatory enzyme, in the synthesis of methionine in seeds, we generated transgenic RNAi seeds with targeted repression of AtCGS during late developmental stages of seeds. Unexpectedly, these seeds accumulated 2.5-fold more methionine than wild-type seeds. To study the nature of these seeds, transcriptomic and primary metabolite profiling were employed using Affymetrix ATH1 microarray and gas chromatography-mass spectrometry analyses, respectively. The results were compared to transgenic Arabidopsis seeds expressing a feedback-insensitive form of AtCGS (named SSE-AtD-CGS) that were previously showed to accumulate up to sixfold more soluble methionine than wild-type seeds. Statistical assessments showed that the nature of transcriptomic and metabolic changes that occurred in RNAi::AtCGS seeds were relatively similar, but to lesser extents, to those previously reported for SSE-AtD-CGS seeds, and linked to the induction of global transcriptomic and metabolic responses associated with stronger desiccation stress. As transgenic seeds obtained by both manipulations exhibited higher, but different methionine levels, the data strongly suggest that these changes depend on the absolute amounts of methionine within seeds and much less to the expression level of AtCGS.

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Higher endogenous methionine in transgenic Arabidopsis seeds affects the composition of storage proteins and lipids

Cited by 17 publications

References 47 publications

Repression of CYSTATHIONINE γ-SYNTHASE in Seeds Recruits the S-Methylmethionine Cycle

Repression of CYSTATHIONINE γ-SYNTHASE in Seeds Recruits the S-Methylmethionine Cycle

In-Depth Investigation of Low-Abundance Proteins in Matured and Filling Stages Seeds of Glycine max Employing a Combination of Protamine Sulfate Precipitation and TMT-Based Quantitative Proteomic Analysis

Dose-dependent effects of higher methionine levels on the transcriptome and metabolome of transgenic Arabidopsis seeds

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